Early life exposure to queen mandibular pheromone mediates persistent transcriptional changes in the brain of honey bee foragers.

Behavioural regulation in insect societies remains a fundamental question in sociobiology. In hymenopteran societies, the queen plays a crucial role in regulating group behaviour by affecting individual behaviour and physiology through modulation of worker gene expression. Honey bee (Apis mellifera) queens signal their presence via queen mandibular pheromone (QMP). While QMP has been shown to influence behaviour and gene expression of young workers, we know little about how these changes translate in older workers. The effects of the queen pheromone could have prolonged molecular impacts on workers that depend on an early sensitive period. We demonstrate that removal of QMP impacts long-term gene expression in the brain and antennae in foragers that were treated early in life (1 day post emergence), but not when treated later in life. Genes important for division of labour, learning, chemosensory perception and ageing were among those differentially expressed in the antennae and brain tissues, suggesting that QMP influences diverse physiological and behavioural processes in workers. Surprisingly, removal of QMP did not have an impact on foraging behaviour. Overall, our study suggests a sensitive period early in the life of workers, where the presence or absence of a queen has potentially life-long effects on transcriptional activity.

Diverse communication strategies in bees as a window into adaptations to an unpredictable world.

Communication is a fundamental feature of animal societies and helps their members to solve the challenges they encounter, from exploiting food sources to fighting enemies or finding a new home. Eusocial bees inhabit a wide range of environments and they have evolved a multitude of communication signals that help them exploit resources in their environment efficiently. We highlight recent advances in our understanding of bee communication strategies and discuss how variation in social biology, such as colony size or nesting habits, and ecological conditions are important drivers of variation in communication strategies. Anthropogenic factors, such as habitat conversion, climate change, or the use of agrochemicals, are changing the world bees inhabit, and it is becoming clear that this affects communication both directly and indirectly, for example by affecting food source availability, social interactions among nestmates, and cognitive functions. Whether and how bees adapt their foraging and communication strategies to these changes represents a new frontier in bee behavioral and conservation research.

Mechanisms and adaptations that shape division of labour in stingless bees.

Stingless bees are a diverse and ecologically important group of pollinators in the tropics. Division of labour allows bee colonies to meet the various demands of their social life, but has been studied in only ∼3% of all described stingless bee species. The available data suggest that division of labour shows both parallels and striking differences compared with other social bees. Worker age is a reliable predictor of worker behaviour in many species, while morphological variation in body size or differences in brain structure are important for specific worker tasks in some species. Stingless bees provide opportunities to confirm general patterns of division of labour, but they also offer prospects to discover and study novel mechanisms underlying the different lifestyles found in eusocial bees.

When should bees be flower constant? An agent-based model highlights the importance of social information and foraging conditions.

Many bee species show flower constancy, that is, a tendency to visit flowers of one type during a foraging trip. Flower constancy is important for plant reproduction, but the benefits of constancy to bees is unclear. Social bees, which often use communication about food sources, show particularly strong flower constancy. We aimed to better understand the benefits of flower constancy in social bees and how these benefits depend on foraging conditions. We hypothesised that sharing social information increases the benefits of flower constancy because social foragers share information selectively about high-quality food sources, thereby reducing the need to sample alternatives. We developed an agent-based model that allowed us to simulate bee colonies with and without communication and flower constancy in different foraging environments. By varying key environmental parameters, such as food source numbers and reward size, we explored how the costs and benefits of flower constancy depend on the foraging landscape. Flower constancy alone performed poorly in all environments, while indiscriminate flower choice was often the most successful strategy. However, communication improved the performance of flower constant colonies considerably in most environments. This combination was particularly successful when high-quality food sources were abundant and competition was weak. Our findings help explain why social bees tend to be more flower constant than solitary bees and suggest that flower constancy can be an adaptive strategy in social bees. Simulations suggest that anthropogenic changes of foraging landscapes will have different effects on the foraging performance of bees that vary in flower constancy.

Sociality is a key driver of foraging ranges in bees.

Bees are important pollinators of wild and agricultural plants1,2,3,4,5 and there is increasing evidence that many bee populations decline due to a combination of habitat loss, climate change, pesticides, and other anthropogenic effects.6,7,8,9,10,11 One trait that shapes both their role in plant reproduction12,13 and their exposure to anthropogenic stressors is the distance at which bees forage. It has been suggested that bee sociality14 and diet15 affect bee foraging ranges, but how these traits and their potential interactions drive foraging ranges remains unclear. We analyzed flight distance data from 90 bee species and developed an agent-based model to test how social, dietary, and environmental factors affect foraging ranges. We confirm that bee sociality is positively associated with foraging range, with average-sized social bees foraging up to 3 times farther from the nest than size-matched solitary bees. A comparative analysis of social bees and computer simulations shows that foraging distances increase with colony size, supporting the hypothesis that greater foraging distances are an emergent property of increasing colony sizes in a food-limited environment. Flower constancy and communication, two traits often found in social bees, synergistically increase foraging distances further in many simulated environments. Diet breadth (oligolectic versus polylectic diet), on the other hand, does not appear to affect foraging ranges in solitary bees. Our findings suggest that multiple traits linked to bee sociality explain why social bees have greater foraging ranges. This has implications for predicting pollination services and for developing effective conservation strategies for bees and isolated plant populations.15,16,17,18,19.

An exploration of the relationship between recruitment communication and foraging in stingless bees.

Social information is widely used in the animal kingdom and can be highly adaptive. In social insects, foragers can use social information to find food, avoid danger, or choose a new nest site. Copying others allows individuals to obtain information without having to sample the environment. When foragers communicate information they will often only advertise high-quality food sources, thereby filtering out less adaptive information. Stingless bees, a large pantropical group of highly eusocial bees, face intense inter- and intra-specific competition for limited resources, yet display disparate foraging strategies. Within the same environment there are species that communicate the location of food resources to nest-mates and species that do not. Our current understanding of why some species communicate foraging sites while others do not is limited. Studying freely foraging colonies of several co-existing stingless bee species in Brazil, we investigated if recruitment to specific food locations is linked to 1) the sugar content of forage, 2) the duration of foraging trips, and 3) the variation in activity of a colony from 1 day to another and the variation in activity in a species over a day. We found that, contrary to our expectations, species with recruitment communication did not return with higher quality forage than species that do not recruit nestmates. Furthermore, foragers from recruiting species did not have shorter foraging trip durations than those from weakly recruiting species. Given the intense inter- and intraspecific competition for resources in these environments, it may be that recruiting species favor food resources that can be monopolized by the colony rather than food sources that offer high-quality rewards.

The adaptive value of tandem communication in ants: Insights from an agent-based model.

Social animals often share information about the location of resources, such as a food source or a new nest-site. One well-studied communication strategy in ants is tandem running, whereby a leader guides a recruit to a resource. Tandem running is considered an example of animal teaching because a leader adjusts her behaviour and invests time to help another ant to learn the location of a resource more efficiently. Tandem running also has costs, such as waiting inside the nest for a leader and a reduced walking speed. Whether and when these costs outweigh the benefits of tandem running is not well understood. We developed an agent-based simulation model to investigate the conditions that favour communication by tandem running during foraging. We predicted that the spatio-temporal distribution of food sources, colony size and the ratio of scouts and recruits affect colony foraging success. Our results suggest that tandem running is favoured when food sources are hard to find, differ in energetic value and are long lasting. These results mirror the findings of simulations of honeybee communication. Scouts locate food sources faster than tandem followers in some environments, suggesting that tandem running may fulfil the criteria of teaching only in some situations. Furthermore, tandem running was only beneficial above a critical colony size threshold. Taken together, our model suggests that there is a considerable parameter range that favours colonies that do not use communication by tandem running, which could explain why many ants with small colony sizes forage solitarily.

Use of waggle dance information in honey bees is linked to gene expression in the antennae, but not in the brain.

Communication is essential for social animals, but deciding how to utilize information provided by conspecifics is a complex process that depends on environmental and intrinsic factors. Honey bees use a unique form of communication, the waggle dance, to inform nestmates about the location of food sources. However, as in many other animals, experienced individuals often ignore this social information and prefer to rely on prior experiences, i.e., private information. The neurosensory factors that drive the decision to use social information are not yet understood. Here we test whether the decision to use social dance information or private information is linked to gene expression differences in different parts of the nervous system. We trained bees to collect food from sugar water feeders and observed whether they utilize social or private information when exposed to dances for a new food source. We performed transcriptome analysis of four brain parts (11-16 bees per tissue type) critical for cognition: the subesophageal ganglion, the central brain, the mushroom bodies, and the antennal lobes but, unexpectedly, detected no differences between social or private information users. In contrast, we found 413 differentially expressed genes in the antennae, suggesting that variation in sensory perception mediates the decision to use social information. Social information users were characterized by the upregulation of biogenic amine genes, while private information users upregulated several genes coding for odour perception. These results highlight that decision-making in honey bees might also depend on peripheral processes of perception rather than higher-order brain centres of information integration.

Forager age and foraging state, but not cumulative foraging activity, affect biogenic amine receptor gene expression in the honeybee mushroom bodies.

Foraging behavior is crucial for the development of a honeybee colony. Biogenic amines are key mediators of learning and the transition from in-hive tasks to foraging. Foragers vary considerably in their behavior, but whether and how this behavioral diversity depends on biogenic amines is not yet well understood. For example, forager age, cumulative foraging activity or foraging state may all be linked to biogenic amine signaling. Furthermore, expression levels may fluctuate depending on daytime. We tested if these intrinsic and extrinsic factors are linked to biogenic amine signaling by quantifying the expression of octopamine, dopamine and tyramine receptor genes in the mushroom bodies, important tissues for learning and memory. We found that older foragers had a significantly higher expression of Amdop1, Amdop2, AmoctαR1, and AmoctßR1 compared to younger foragers, whereas Amtar1 showed the opposite pattern. Surprisingly, our measures of cumulative foraging activity were not related to the expression of the same receptor genes in the mushroom bodies. Furthermore, we trained foragers to collect sucrose solution at a specific time of day and tested if the foraging state of time-trained foragers affected receptor gene expression. Bees engaged in foraging had a higher expression of Amdop1 and AmoctßR3/4 than inactive foragers. Finally, the expression of Amdop1, Amdop3, AmoctαR1, and Amtar1 also varied with daytime. Our results show that receptor gene expression in forager mushroom bodies is complex and depends on both intrinsic and extrinsic factors.

Correlation between octopaminergic signalling and foraging task specialisation in honeybees.

Regulation of pollen and nectar foraging in honeybees is linked to differences in the sensitivity to the reward. Octopamine (OA) participates in the processing of reward-related information in the bee brain, being a candidate to mediate and modulate the division of labour among pollen and nectar foragers. Here we tested the hypothesis that OA affects the resource preferences of foragers. We first investigated whether oral administration of OA is involved in the transition from nectar to pollen foraging. We quantified the percentage of OA-treated bees that switched from a sucrose solution to a pollen feeder when the sugar concentration was decreased experimentally. We also evaluated if feeding the colonies sucrose solution containing OA increases the rate of bees collecting pollen. Finally, we quantified OA and tyramine (TYR) receptor genes expression of pollen and nectar foragers in different parts of the brain, as a putative mechanism that affects the decision-making process regarding the resource type collected. Adding OA in the food modified the probability that foragers switch from nectar to pollen collection. The proportion of pollen foragers also increased after feeding colonies with OA-containing food. Furthermore, the expression level of the AmoctαR1 was upregulated in foragers arriving at pollen sources compared with those arriving at sugar-water feeders. Using age-matched pollen and nectar foragers that returned to the hive, we detected an upregulated expression of a TYR receptor gene in the suboesophageal ganglia. These findings support our prediction that OA signalling affects the decision in honeybee foragers to collect pollen or nectar.

Octopamine and dopamine mediate waggle dance following and information use in honeybees.

Honeybees can be directed to profitable food sources by following waggle dances performed by other bees. Followers can often choose between using this social information or relying on memories about food sources they have visited in the past, so-called private information. While the circumstances that favour the use of either social or private information have received considerable attention, still little is known about the neurophysiological basis of information use. We hypothesized that octopamine and dopamine, two biogenic amines with important functions in reward signalling and learning, affect dance use in honeybees. We orally administered octopamine and dopamine when bees collected food at artificial feeders and tested if this affected interest in dance information about a new food source. We predicted that octopamine reduces interest in dances and strengthens private information use via an increase in the perceived value of the previously exploited resource. Since dopamine has been shown to lower reward perception, we expected it to act in the opposite direction. Octopamine-treated foragers indeed followed 32% fewer dances than control bees and increased the use of private information. Conversely, dopamine-treated bees followed dances 15% longer than control bees, but surprisingly did not use social information more. Overall, our results suggest that biogenic amine signalling affects interactions among dancers and dance followers and, thus, information flow about high-quality food sources.

Octopamine increases individual and collective foraging in a neotropical stingless bee.

The biogenic amine octopamine (OA) is a key modulator of individual and social behaviours in honeybees, but its role in the other group of highly eusocial bees, the stingless bees, remains largely unknown. In honeybees, OA mediates reward perception and affects a wide range of reward-seeking behaviours. Thus, we tested the hypothesis that OA increases individual foraging effort and collective food source exploitation in the neotropical stingless bee Plebeia droryana. OA treatment caused a significant increase in the number of bees at artificial sucrose feeders and a 1.73-times higher individual foraging frequency. This effect can be explained by OA lowering the sucrose response threshold and, thus, increasing the perceived value of the food source. Our results demonstrate that, similar to its effects on honeybees, OA increases both individual and collective food source exploitation in P. droryana. This suggests that, despite having evolved many complex behaviours independently, OA might have similar regulatory effects on foraging behaviours in the two groups of highly eusocial bees.

Resource profitability, but not caffeine, affects individual and collective foraging in the stingless bee Plebeia droryana.

Plants and pollinators form beneficial relationships, with plants offering resources in return for pollination services. Some plants, however, add compounds to nectar to manipulate pollinators. Caffeine is a secondary plant metabolite found in some nectars that affects foraging in pollinators. In honeybees, caffeine increases foraging and recruitment to mediocre food sources, which might benefit the plant, but potentially harms the colonies. For the largest group of social bees, the stingless bees, the effect of caffeine on foraging behaviour has not been tested yet, despite their importance for tropical ecosystems. More generally, recruitment and foraging dynamics are not well understood in most species. We examined whether caffeine affects the foraging behaviour of the stingless bee Plebeia droryana, which frequently visits plants that produce caffeinated nectar and pollen. We trained bees to food sources containing field-realistic concentrations of sugar and caffeine. Caffeine did not cause P. droryana to increase foraging frequency and persistence. We observed P. droryana recruiting to food sources; however, this behaviour was also not affected by caffeine. Instead we found that higher sugar concentrations caused bees to increase foraging effort. Thus, unlike in other pollinators, foraging behaviour in this stingless bee is not affected by caffeine. As the Brazilian P. droryana population that we tested has been exposed to coffee over evolutionary time periods, our results raise the possibility that it may have evolved a tolerance towards this central nervous system stimulant. Alternatively, stingless bees may show physiological responses to caffeine that differ from those of other bee groups.

Repeated switches from cooperative to selfish worker oviposition during stingless bee evolution.

Reproductive division of labour is a defining feature of insect societies. Stingless bees (Meliponini) are an interesting exception among the highly eusocial insects in that workers of many species contribute significantly to the production of males. Since workers remain sterile in other species of this large tropical tribe, it has been hypothesized that, in the latter species, ancestral queens have won the conflict over who produces the males. The fact that sterile workers of some species lay trophic eggs to feed the queen and display ritualized behaviours towards her during oviposition has been interpreted as an evolutionary relic of this ancient conflict. Here, I used ancestral state estimation to test whether worker reproduction is indeed the ancestral condition and worker sterility a derived state in stingless bees. Contrary to this hypothesis, data suggest that trophic egg laying was the ancestral condition, whereas selfish worker reproduction in queenright colonies evolved subsequently during stingless bee diversification. The appearance of worker reproduction in queenright conditions was tightly linked to the laying of trophic eggs, which suggests that having activated ovaries in queen presence facilitates the evolution of worker reproduction. Worker reproduction is also linked to brood cell architecture, but surprisingly not to colony size or queen-worker dimorphism. The reason for this association between brood cell architecture and worker oviposition is currently unknown. These results suggest that trophic eggs are not a relic of an ancient conflict, but a sign of overlapping interests between the queen and workers about who produces the males.

Insect societies fight back: the evolution of defensive traits against social parasites.

Insect societies face many social parasites that exploit their altruistic behaviours or their resources. Due to the fitness costs these social parasites incur, hosts have evolved various behavioural, chemical, architectural and morphological defence traits. Similar to bacteria infecting multicellular hosts, social parasites have to successfully go through several steps to exploit their hosts. Here, we review how social insects try to interrupt this sequence of events. They can avoid parasite contact by choosing to nest in parasite-free locales or evade attacks by adapting their colony structure. Once social parasites attack, hosts attempt to detect them, which can be facilitated by adjustments in colony odour. If social parasites enter the nest, hosts can either aggressively defend their colony or take their young and flee. Nest structures are often shaped to prevent social parasite invasion or to safeguard host resources. Finally, if social parasites successfully establish themselves in host nests, hosts can rebel by killing the parasite brood or by reproducing in the parasites' presence. Hosts of social parasites can therefore develop multiple traits, leading to the evolution of complex defence portfolios of co-dependent traits. Social parasites can respond to these multi-level defences with counter-adaptations, potentially leading to geographical mosaics of coevolution.This article is part of the Theo Murphy meeting issue 'Evolution of pathogen and parasite avoidance behaviours'.

Enemy recognition is linked to soldier size in a polymorphic stingless bee.

Many ant and termite colonies are defended by soldiers with powerful mandibles or chemical weaponry. Recently, it was reported that several stingless bee species also have soldiers for colony defence. These soldiers are larger than foragers, but otherwise lack obvious morphological adaptations for defence. Thus, how these soldiers improve colony fitness is not well understood. Robbing is common in stingless bees and we hypothesized that increased body size improves the ability to recognize intruders based on chemosensory cues. We studied the Neotropical species Tetragonisca angustula and found that large soldiers were better than small soldiers at recognizing potential intruders. Larger soldiers also had more olfactory pore plates on their antennae, which is likely to increase their chemosensory sensitivity. Our results suggest that improved enemy recognition might select for increased guard size in stingless bees.

Repeated evolution of soldier sub-castes suggests parasitism drives social complexity in stingless bees.

The differentiation of workers into morphological castes represents an important evolutionary innovation that is thought to improve division of labor in insect societies. Given the potential benefits of task-related worker differentiation, it is puzzling that physical worker castes, such as soldiers, are extremely rare in social bees and absent in wasps. Following the recent discovery of soldiers in a stingless bee, we studied the occurrence of worker differentiation in 28 stingless bee species from Brazil and found that several species have specialized soldiers for colony defence. Our results reveal that worker differentiation evolved repeatedly during the last ~ 25 million years and coincided with the emergence of parasitic robber bees, a major threat to many stingless bee species. Furthermore, our data suggest that these robbers are a driving force behind the evolution of worker differentiation as targets of robber bees are four times more likely to have nest guards of increased size than non-targets. These findings reveal unexpected diversity in the social organization of stingless bees.Although common in ants and termites, worker differentiation into physical castes is rare in social bees and unknown in wasps. Here, Grüter and colleagues find a guard caste in ten species of stingless bees and show that the evolution of the guard caste is associated with parasitization by robber bees.

Soldiers in a Stingless Bee.

The differentiation of workers into morphological subcastes (e.g., soldiers) represents an important evolutionary transition and is thought to improve division of labor in social insects. Soldiers occur in many ant and termite species, where they make up a small proportion of the workforce. A common assumption of worker caste evolution is that soldiers are behavioral specialists. Here, we report the first test of the "rare specialist" hypothesis in a eusocial bee. Colonies of the stingless bee Tetragonisca angustula are defended by a small group of morphologically differentiated soldiers. Contrary to the rare specialist hypothesis, we found that soldiers worked more (+34%-41%) and performed a greater variety of tasks (+23%-34%) than other workers, particularly early in life. Our results suggest a "rare elite" function of soldiers in T. angustula, that is, that they perform a disproportionately large amount of the work. Division of labor was based on a combination of temporal and physical castes, but soldiers transitioned faster from one task to the next. We discuss why the rare specialist assumption might not hold in species with a moderate degree of worker differentiation.

Local differences in parasitism and competition shape defensive investment in a polymorphic eusocial bee.

Many colonial animals rely for their defense on a soldier caste. Adaptive colony demography theory predicts that colonies should flexibly adjust the investment in different worker castes depending on the colony needs. For example, colonies should invest more in defensive workers (e.g., soldiers) in dangerous environments. However, evidence for this prediction has been mixed. We combined descriptive and experimental approaches to examine whether defensive investment and worker size are adjusted to local ecology in the only known bee with polymorphic workers, Tetragonisca angustula. Colonies of this species are defended by a morphologically specialized soldier caste. Our study included three populations that differed in the density of food competition and the occurrence of a parasitic robber bee. We found that colonies coexisting with robber bees had on average 43% more soldiers defending the nest entrance, while colonies facing stronger foraging competition had soldiers that were -6-7% smaller. We then experimentally relocated colonies to areas with different levels of competition. When released from intense food competition, body sizes of guards and foragers increased. After introducing chemical robber bee cues at nest entrances, we found both a short-term and a long-term up-regulation of the number of soldiers defending the colony. Active soldier numbers remained high after the experiment for a duration equivalent to 2-3 worker life spans. How information about past parasite threat is stored in the colony is currently unknown. In summary, T. angustula adjusts both the number and the body size of active soldiers to local ecological conditions. Competitor density also affects forager (or minor) size, an important colony trait with potential community ecological consequences. Our study supports adaptive colony demography theory in a eusocial bee and highlights the importance of colony threats and competition as selective forces shaping colony phenotype.

Inter-caste communication in social insects.

Social insect colonies function as highly integrated units despite consisting of many individuals. This requires the different functional parts of the colony (e.g. different castes) to exchange information that aid in colony functioning and ontogeny. Here we discuss inter-caste communication in three contexts, firstly, the communication between males and females during courtship, secondly, the communication between queens and workers that regulate reproduction and thirdly, the communication between worker castes that allows colonies to balance the number of different worker types. Some signals show surprising complexity in both their chemistry and function, whereas others are simple compounds that were probably already used as pheromones in the solitary ancestors of several social insect lineages.