The effect of queen-worker conflict on caste determination in the bumblebee Bombus terrestris

Endocrine analyses were used to investigate the well-known association between queen production and the onset of worker reproduction (termed the competition phase, CPh) in Bombus terrestris. Larvae that reached the age of 5 days before the CPh had a worker-like profile: low juvenile hormone (JH) biosynthesis rates and low JH hemolymph titers. In contrast, larvae that reached the age of 5 days during the CPh had a queen-like profile: high JH biosynthesis rates and high hemolymph JH levels. Larval fate could be manipulated by transplanting egg cells into host colonies with different social structures. There was a steep rise in JH production in larvae transplanted into colonies near or during the CPh. This indicates that during colony development, larvae switch from the ”worker developmental pathway” to the ”queen developmental pathway,” and that the switch is socially regulated. In small rearing groups, larvae reared with queens before the CPh developed into workers, whereas those reared with queens after the CPh developed into queens. Variation in worker type (naive or experienced) did not affect caste determination. Therefore, we hypothesize that queens produce a pheromone that directly inhibits queen differentiation by larvae. We also present two alternative scenarios that explain the timing of gyne production in B. terrestris, one based on ecological constraints and the other based on queen-worker competition. Received: 20 October 1999 / Received in revised form: 18 December 1999 / Accepted: 23 January 2000     

A comparison of visual and olfactory learning performance in the bumblebee Bombus terrestris

Animals use cues from a range of sensory modalities to discriminate stimuli and as predictors of reward. Whilst there is appreciable variation in the cognitive performance of animals, we know surprisingly little about the extent to which learning varies among individuals across different sensory modalities. Do individuals that are good at learning in one sensory modality also perform well in another (performance is correlated between modalities), or do individuals demonstrate specialisation in learning performance in one modality (trading-off performance between modalities)? We tested these hypotheses by examining the performance of 76 Bombus terrestris workers, from four colonies, in both an odour-and visual learning task. Olfactory learning was assessed using proboscis extension reflex (PER) conditioning and visual (colour) learning was examined using a well-established free-flying paradigm. Our results showed neither a correlation, nor a trade-off, in individual performance for learning tasks using different sensory modalities. However, there was considerable variation among workers within each colony in their performance in both learning tasks. This extent of interindividual variation in learning ability across sensory modalities could be adaptive for colonies dealing with changeable foraging conditions. There was also significant intercolony variation in final task performance level in the olfactory learning task, and both the strength and persistence of blue preference in the colour learning task. This is the first study to demonstrate variation in olfactory learning performance across multiple bumblebee colonies using PER conditioning, suggesting this is an effective paradigm for assessing associative olfactory learning performance both within and among colonies.     

Regulation of reproduction by dominant workers in bumblebee (Bombus terrestris) queenright colonies

The mechanisms of regulating worker reproduction in bumblebees were studied by direct behavioral observations and by measuring ovarian development and juvenile hormone (JH) biosynthesis rates in workers under different social conditions. Workers in the last stage of Bombus terrestris colony development (the competition phase) had the lowest ovarian development and JH biosynthesis rates. Callows introduced into colonies immediately after queen removal (dequeened colonies) demonstrated a significant increase in ovarian development before, but not during, the competition phase. These findings differ from the higher ovarian development in colonies during the competition phase predicted by the prevailing hypothesis that worker reproduction starts in response to a decrease in queen inhibition. Reproduction of callows housed with dominant workers in small queenless groups was inhibited as in queenright colonies. This suggests that the reduced ovarian development and JH biosynthesis rates observed in dequeened and normally developing colonies during the competition phase also reflect inhibition by dominant workers. Thus, two distinct stages of inhibition of reproduction seem to exist: (1) before the competition phase, when the queen slows down worker ovarian development and prevents oviposition; (2) during the competition phase, when dominant workers inhibit ovarian development of other workers. Between these stages there seems to be a temporal “window” of enhanced worker reproductive development. The queen's typical switch to haploid egg production was not associated with changes in worker ovarian development or JH biosynthesis rates. These findings suggest that regulation of worker reproduction in B. terrestris is not determined by simple changes in the queen's inhibition capacity or by the sex of offspring and that the worker's role is more important than previously believed. Received: 18 March 1998 / Accepted after revision: 18 July 1998     

Colony nutritional status modulates worker responses to foraging recruitment pheromone in the bumblebee Bombus terrestris

Foraging activity in social insects should be regulated by colony nutritional status and food availability, such that both the emission of, and response to, recruitment signals depend on current conditions. Using fully automatic radio-frequency identification (RFID) technology to follow the foraging activity of tagged bumblebees (Bombus terrestris) during 16,000 foraging bouts, we tested whether the cue provided by stored food (the number of full honeypots) could modulate the response of workers to the recruitment pheromone signal. Artificial foraging pheromones were applied to colonies with varied levels of food reserves. The response to recruitment pheromones was stronger in colonies with low food, resulting in more workers becoming active and more foraging bouts being performed. In addition to previous reports showing that in colonies with low food successful foragers perform more excited runs during which they release recruitment pheromone and inactive workers are more prone to leave the nest following nectar influx, our results indicate that evolution has shaped a third pathway that modulates bumblebee foraging activity, thus preventing needless energy expenditure and exposure to risk when food stores are already high. This new feedback loop is intriguing since it involves context-dependent response to a signal. It highlights the integration of information from both forager-released pheromones (signal) and nutritional status (cue) that occurs within individual workers before making the decision to start foraging. Our results support the emerging view that responses to pheromones may be less hardwired than commonly acknowledged. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Role of juvenile hormone biosynthesis in dominance status and reproduction of the bumblebee,Bombus terrestris

Summary In Bombus terrestris females, dominant status is correlated with high levels of juvenile hormone (JH) biosynthesis and rapid oöcyte maturation. Queenright workers, which are inhibited by the dominant queen, complete the cycle of oöcyte maturation while exhibiting a continuous low rate of JH production, but their egglaying is inhibited. Measurements in foundress queens suggest that the low JH titer during oögenesis is probably not responsible for the inhibition of egg-laying. Queenless workers, kept individually, are not activated either for JH production or oöcyte maturation. In groups of three queenless workers, a dominance order becomes established and high rates of JH synthesis are observed in the dominant egg-laying workers, with low rates in subordinated workers. In groups of founder queens, also, a dominance order becomes established and results in a reduced rate of JH production in subordinated females. Correspondence to: F. Couillaud     

Effects of experience and weather on foraging rate and pollen versus nectar collection in the bumblebee, Bombus terrestris

This study examines factors that affect foraging rate of free-flying bumblebees, Bombus terrestris, when collecting nectar, and also what factors determine whether they collect pollen or nectar. We show that nectar foraging rate (mass gathered per unit time) is positively correlated with worker size, in accordance with previous studies. It has been suggested that the greater foraging rate of large bees is due to their higher thermoregulatory capacity in cool conditions, but our data suggest that this is not so. Workers differing in size were not differentially affected by the weather. Regardless of size, naïve bees were poor foragers, often using more resources than they gathered. Foraging rate was not maximised until at least 30 trips had been made from the nest. Foraging rates were positively correlated with humidity, perhaps because nectar secretion rates were higher or evaporation of nectar lower at high humidity. Temperature, wind speed and cloud cover did not significantly influence foraging rate, within the summertime range that occurred during the study. Weather greatly influenced whether bees collected pollen or nectar. Pollen was preferably collected when it was warm, windy, and particularly when humidity was low; and preferably during the middle of the day. We suggest that bees collect pollen in dry conditions, and avoid collecting pollen when there is dew or rain-water droplets on the vegetation, which would make grooming pollen into the corbiculae difficult. Availability of sufficient dry days for pollen collection may be an important factor determining the success of bumblebee colonies.Communicated by M. Giurfa     

Reproductive decision-making in semelparous colonies of the bumblebee bombus terrestris

Queen and worker Bombus terrestris have different optima for the timing of gyne production. Workers, being more related to their gyne-sisters than to their sons, should ascertain that gyne production has started before attempting to reproduce. Their optimal timing for gyne production will be as early as possible, while allowing sufficient ergonomic colony growth to support gyne rearing. Queen optimum, on the other hand, should be to postpone gyne production toward the end of colony life cycle, in order to minimize the time-window available for worker reproduction. Thus, the timing of gyne production may profoundly affect the outcome of queen–worker competition over male production. In this study we investigated some of the social correlates possibly affecting this timing. It was found that neither keeping colony size constant and as low as 20 workers, nor decreasing worker average age, influenced the onset of gyne production. To test the effect of queen age we created young colonies with old queens and vice versa. When colony social composition remained unchanged, in young colonies headed by old queens gynes were produced earlier than predicted, but in the inverse situation gyne production was not delayed. When colony social composition was completely standardized queen age had a decisive effect, indicating that the timing of gyne production is both under queen influence and affected by queen age. Furthermore, queens assess colony age from the time of first worker emergence rather than from their own first oviposition. In these experiments the factors affecting gyne production also affected the onset of queen–worker conflict for male production, suggesting that both are regulated by the same causal effect. Postponing gyne production as much as possible provides another mechanism, in addition to extensive oophagy, for the queen to outcompete her workers in male production.     

Worker drift and egg dumping by queens in wild Bombus terrestris colonies

Wild bumblebee colonies are hard to find and often inaccessible, so there have been few studies of the genetic structure of bumblebees within natural colonies, and hence, it is not clear how frequently events such as worker reproduction, worker drift and queen usurpation take place. This study aimed to quantify the occurrence of natal-worker reproduction, worker drift and drifter reproduction within 14 wild colonies of Bombus terrestris in Central Scotland. Four unlinked microsatellites were used to identify patterns of relatedness of the colonies’ adults and broods. In colonies with queens (queenright colonies), worker reproduction accounted for just 0.83 % of males, increasing to 12.11 % in queenless colonies. Four colonies contained a total of six workers which were not daughters of the queen, and were assumed to be drifters, and four male offspring of drifters. Drifting is clearly not common and results in few drifter offspring overall, although drifters produced approximately seven times more offspring per capita than workers that remained in their natal colony. Unexpectedly, two colonies contained clusters of sister workers and juvenile offspring that were not sisters to the rest of the adults or brood found in the colonies, demonstrating probable egg dumping by queens. A third colony contained a queen which was not a sister or daughter to the other bees in the colony. Although usurping of bumblebee colonies by queens in early season is well documented, this appears to be the first record of egg dumping, and it remains unclear whether it is being carried out by old queens or newly mated young queens.     

Food alert in bumblebees (Bombus terrestris): possible mechanisms and evolutionary implications

The return of a successful bumblebee forager stimulates nestmates to leave the nest and search for food. Here we investigate the mechanisms by which this happens. Successful Bombus terrestris foragers perform irregular runs in their nest, often lasting for several minutes. Run duration is at its maximum when food has just been discovered. Running likely serves to distribute a pheromone, since the information flow between "runners" and "recruits" can be disrupted by eliminating air exchange, while leaving other potential means of communication intact. In addition, nectar stores in the nest may be monitored continuously. A sudden influx of nectar into the nest also causes measurable increases in forager activity. The implications of bumblebee recruitment behavior for the evolution of communication in bees are discussed.     

The appeasement effect of sterility signaling in dominance contests among Bombus terrestris workers

The establishment of dominance hierarchies through aggressive interactions is very common in insect societies. In many cases, it is also mediated through pheromone emissions that enable individuals to evaluate the reproductive quality and level of aggressiveness of the dominant individual, thereby reducing the number and intensity of costly fights. Here, we studied these processes in the primitively eusocial bee Bombus terrestris, using a paired bee system. Specifically, we investigated the behavioral, reproductive, and pheromonal correlates of dominance establishment. Workers were shown to establish dominance hierarchies using overt aggression within 3–4 days. Thereafter, the aggression drastically decreased, and dominance was maintained mostly by ritualized agonistic behavior. The behaviorally dominant bee lost the ester compounds that workers produce in their Dufour's gland (the so-called “sterility signal”) concomitantly with the development of her ovaries. The other bee announced as subordinate by continuously producing high amounts of those esters. The hypothesis that sterility signaling serves as an appeasement signal to pacify the dominant bees is supported by the negative correlation found between the proportion of these esters and the level of aggression that the subordinate received from the dominant worker. Physical interactions, and presumably also the ensuing overt aggression between the bees, were essential for the above pheromonal change to take place and enabled the dominant workers to develop their ovaries and to lay eggs. The subordinate bee’s signaling of non-reproductive status may minimize energy expenditure in costly fights and help stabilize the reproductive division of labor among workers.     

Seasonal activity of Bombus terrestris L. in east Mediterranean region, Turkey

Bumblebees have economical importance in most of wild and cultivated plants. They can be abundant in suitable habitats and have a broad flower choice. Bombus terrestris was collected at intervals during 2002 and 2003 from various flora and ecosystems of east Mediterranean region of Turkey. In this study, plants visited by Bombus terrestris, seasonal activities, distribution and altitudes were determined. Bombus terrestris have boon seen throughout Turkey in a wide range of habitats from sea level to 1560 m altitude within all the major native vegetation types. Prevalence of queens, workers and males of Bombus terrestris differed due to altitude. More frequently observed at 0-600 m, declining above 600 m in relation to general climate requirements.     

Piercing the partner's skin influences sperm uptake in the earthworm Lumbricus terrestris

Sexual conflict between mating partners can give rise to strategies that are advantageous for one sex but harmful to the opposite sex. Usually, sperm donors develop (offensive) traits to enhance their chances in sperm competition, while sperm recipients evolve (defensive) traits that allow them to stay in control of who fathers their offspring. Here, we demonstrate that these processes are also at work in simultaneous hermaphrodites. The hermaphroditic earthworm Lumbricus terrestris uses 40 to 44 copulatory setae to pierce into its partner's skin, causing damage and injecting a substance from its setal glands. Experimental injection of the gland substance indicates that a refractory period may be induced. More importantly, removal of the copulatory setae shows that they influence the partner's sperm uptake. When the setae are present, more sperm are taken up and sperm are distributed more equally over the four spermathecae. We interpret this as a strategy that stacks the odds for the donor's sperm in fertilizing cocoons.     

Regulation of worker reproduction in bumblebees (Bombus terrestris): workers eavesdrop on a queen signal

In the annual bumblebee Bombus terrestris, the onset of queen-worker conflict over male production is seasonally and socially constrained. Workers will do better if they start to reproduce (the so-called competition phase) only after ascertaining that larvae are committed to gyne development but before the season ends because they gain more by rearing sister-gynes than their own sons. Here, we tested two nonmutually exclusive hypotheses as to what triggers the onset of worker reproduction: Workers can directly monitor larval development and/or workers eavesdrop on the queen signal that directs gyne development. Exposing workers to gyne larvae through a double mesh did not advance the competition phase compared to control colonies. However, when workers, but not the queen, were allowed contact with gyne larvae, both the competition phase and gyne production were advanced. Thus, while larvae do not emit a volatile pheromone that discloses their developmental route, the physical contact of workers with such larvae triggers early competition phase. However, workers exclusively exposed to worker larvae (colonies prevented from producing gyne larvae) started to reproduce at the same time as control colonies. Replacing the resident queen with an older queen (from gyne-rearing colonies) advanced the competition phase, irrespective of worker age. The results are consistent with the hypothesis that workers eavesdrop on the queen pheromones. This is adaptive because it allows workers a broader time-window for reproduction and thus to gain fitness from rearing both sister-gynes and sons before the season ends without affecting colony development.     

Transmission of a pathogen in Bombus terrestris,with a note on division of labour in social insects

Summary Parasites of social insect workers can be transmitted within the colony to other, related host individuals or, alternatively, to unrelated workers of other colonies. Division of labour affects the probability of transmission, as young individuals often work inside the nest whereas older ones often leave the nest to forage. Therefore, the relative probabilities of transmission within-vs. between-nests is also affected by the delay between host infection and the shedding of propagules, i.e. the latent period of the parasite strain. We therefore hypothesized that strains of the flagellate parasite Crithidia bombi (Trypanosomatidae, Zoomastigophorea) infecting workers of the bumble bee Bombus terrestris (Hymenoptera, Apidae) could differ in their delays and coexist in a population. This would be the case if strains that are shed after a short time delay were more efficiently transmitted to other colony members, whereas strains with long delays were more efficiently transmitted to non-related workers in the population. We tested this hypothesis by experimentally varying time delay and by allowing transmission to either sister workers from the same nest or unrelated workers from other nests. Transmission of C. bombi was measured as the number of parasitic cells shed by the exposed workers after a standard period. The results showed that relatedness as such had no effect, but that delay and nest identity were highly significant effects to explain variation in transmission success. There was a significant interaction between nest identity and delay, such that bees of some colonies acted as efficient transmitters for C. bombi under short delays and vice versa. We discuss how division of labour may affect parasitism in social insects and, vice versa, how division of labour may be under selection from the effects of parasitism, using available evidence from the literature. Correspondence to: P. Schmid-Hempel     

Color choices by bumble bees (Bombus terrestris): innate preferences and generalization after learning

It is usually assumed that the choice behavior of bees for floral colors is influenced by innate preferences only for the first flower visits prior to any experience. After visits to rewarding flowers bees learn to associate their colors with a reward. This learning process leads to an acquired preference for the trained colors that has been believed to dominate over previous experiences and over innate preferences. This work investigates how bumble bees (Bombus terrestris) chose among artificial flowers of different colors after they had been extensively trained to other colors. The bees chose novel colors according to their similarity to the trained color if the trained color was similar to some of the test colors. This was true also if trained colors and test colors were well distinguished, so their color choice reflected generalization between colors. If the test colors were so different from the trained color that no generalization took place, choice behavior was not affected by the trained color and reflected innate preferences. The differences in choice frequencies could not be explained by physical properties of the test colors other than the dominant wavelength, a parameter taken to reflect hue perception. Preferred dominant wavelengths correspond to those observed in naive bumble bees and honeybees. Thus bumble bees show innate preferences for certain colors not only prior to color learning but also after intensive learning when choosing among very different novel colors. Color choice among similar colors, however, is controlled by generalization from the learned color. Received: 9 November 1999 / Received in revised form: 19 March 2000 / Accepted: 31 March 2000     

Social influences on the whistled songs of starlings

Summary Playback experiments on individuals singing near their nest show that starlings tend to repeat the theme they have just heard even if it is an unfamiliar dialect and not programmed in their own current sequence. The type of response and its temporal characteristics could be related to internal sequential organization rules. Moreover, starlings discriminate between two dialectal variants of a given theme and respond more often and more quickly to the familiar dialect. A social influence on the choice of theme by the individual can also be observed in foraging flocks and roosts where song is frequently given. Observation of such groups reveal that one theme predominates on each given occasion and that it depends on the number of individuals present and on the type of group (flock or roost). This is probably a function of the tendency to match. Previous observations of flights toward the roost showed that birds originating from different dialectal areas congregate in such groups. A few analyses of songs in flocks and roosts confirm the presence of different dialects simultaneously.     

Two phylogenetically distinct species of sexually deceptive orchids mimic the sex pheromone of their single common pollinator, the cuckoo bumblebee Bombus vestalis

Sexually deceptive orchids of the genus Ophrys attract male insects for pollination. Pollinator attraction is achieved by mimicking sex pheromones of virgin females of their pollinators, mostly bee species. In earlier investigations, we showed that the phylogenetically distinct Ophrys species O. chestermanii and O. normanii on Sardinia attract their pollinator, males of the cuckoo bumblebee B. vestalis, with the same bouquets of relatively polar volatile compounds. In this investigation, we studied the sex pheromone of virgin females of B. vestalis with the aim of identifying male-attracting compounds and of comparing them with labellum extracts of the two orchids, which were found to release male-attracting compounds in earlier investigations (G?gler et al. 2009). In bioassays, shock-frozen females, cuticle extracts and polar fractions of cuticle extracts of virgin females stimulated mating behaviour in the males. Using gas chromatography coupled with electroantennography (GC-EAD) and gas chromatography coupled with mass spectrometry (GC-MS), we detected in polar fractions of cuticle extracts of B. vestalis females the same electrophysiologically active compounds as in labellum extracts of both orchid species, including aldehydes, esters, fatty acids and alcohols. Since statistical comparisons of the relative proportions of esters showed strong similarities between virgin females and orchids, our results support the hypotheses that this highly specialized Ophrys–pollinator relationship represents another case of chemical mimicry and that esters play a key role in male attraction.