Signatures of adaptive decreased virulence of deformed wing virus in an isolated population of wild honeybees (Apis mellifera).

Understanding the ecological and evolutionary processes that drive host-pathogen interactions is critical for combating epidemics and conserving species. The Varroa destructor mite and deformed wing virus (DWV) are two synergistic threats to Western honeybee (Apis mellifera) populations across the globe. Distinct honeybee populations have been found to self-sustain despite Varroa infestations, including colonies within the Arnot Forest outside Ithaca, NY, USA. We hypothesized that in these bee populations, DWV has been selected to produce an avirulent infection phenotype, allowing for the persistence of both host and disease-causing agents. To investigate this, we assessed the titre of viruses in bees from the Arnot Forest and managed apiaries, and assessed genomic variation and virulence differences between DWV isolates. Across groups, we found viral abundance was similar, but DWV genotypes were distinct. We also found that infections with isolates from the Arnot Forest resulted in higher survival and lower rates of symptomatic deformed wings, compared to analogous isolates from managed colonies, providing preliminary evidence to support the hypothesis of adaptive decreased viral virulence. Overall, this multi-level investigation of virus genotype and phenotype indicates that host ecological context can be a significant driver of viral evolution and host-pathogen interactions in honeybees.

Remembrances of a Honey Bee Biologist.

Thomas Seeley's research has focused on analyzing the collective intelligence and natural lives of honey bees. This account describes how the author encountered honey bees as a boy and became a beekeeper; how he switched his career path from medicine to biology to study the behavior and social life of honey bees; and how he focuses on understanding how a honey bee colony functions when it lives in the wild, rather than in a beekeeper's hive. He has shown how a honey bee colony works as a single decision-making unit to adaptively allocate its foragers among flower patches and to choose its nesting site in a hollow tree. These findings buttress the view that, in some social insect species, the colony is a group-level vehicle of gene survival. Beyond his research, he has written three books to synthesize these findings for biologists and share these discoveries with beekeepers.

Varroa destructor Mites Can Nimbly Climb from Flowers onto Foraging Honey Bees.

Varroa destructor, the introduced parasite of European honey bees associated with massive colony deaths, spreads readily through populations of honey bee colonies, both managed colonies living crowded together in apiaries and wild colonies living widely dispersed in natural settings. Mites are hypothesized to spread between most managed colonies via phoretically riding forager bees when they engage in robbing colonies or they drift between hives. However, widely spaced wild colonies show Varroa infestation despite limited opportunities for robbing and little or no drifting of bees between colonies. Both wild and managed colonies may also exchange mites via another mechanism that has received remarkably little attention or study: floral transmission. The present study tested the ability of mites to infest foragers at feeders or flowers. We show that Varroa destructor mites are highly capable of phoretically infesting foraging honey bees, detail the mechanisms and maneuvers by which they do so, and describe mite behaviors post-infestation.

The behavioral regulation of thirst, water collection and water storage in honey bee colonies.

This study investigated how a honey bee colony develops and quenches its collective thirst when it experiences hyperthermia of its broodnest. We found that a colony must strongly boost its water intake because evaporative cooling is critical to relieving broodnest hyperthermia, and that it must rapidly boost its water intake because a colony maintains only a small water reserve. We also clarified how a colony's water collectors know when to spring into action - by sensing either more frequent requests for fluid or greater personal thirst, or both. Finally, we found that the behavioral flexibility of a colony's water collectors enables them not only to satisfy their colony's current water needs but also to buffer their colony against future extreme water stresses by storing water in their crops and in their combs.

How Honey Bee Colonies Survive in the Wild: Testing the Importance of Small Nests and Frequent Swarming.

The ectoparasitic mite, Varroa destructor, and the viruses that it transmits, kill the colonies of European honey bees (Apis mellifera) kept by beekeepers unless the bees are treated with miticides. Nevertheless, there exist populations of wild colonies of European honey bees that are persisting without being treated with miticides. We hypothesized that the persistence of these wild colonies is due in part to their habits of nesting in small cavities and swarming frequently. We tested this hypothesis by establishing two groups of colonies living either in small hives (42 L) without swarm-control treatments or in large hives (up to 168 L) with swarm-control treatments. We followed the colonies for two years and compared the two groups with respect to swarming frequency, Varroa infesttion rate, disease incidence, and colony survival. Colonies in small hives swarmed more often, had lower Varroa infestation rates, had less disease, and had higher survival compared to colonies in large hives. These results indicate that the smaller nest cavities and more frequent swarming of wild colonies contribute to their persistence without mite treatments.

Museum samples reveal rapid evolution by wild honey bees exposed to a novel parasite.

Understanding genetic changes caused by novel pathogens and parasites can reveal mechanisms of adaptation and genetic robustness. Using whole-genome sequencing of museum and modern specimens, we describe the genomic changes in a wild population of honey bees in North America following the introduction of the ectoparasitic mite, Varroa destructor. Even though colony density in the study population is the same today as in the past, a major loss of haplotypic diversity occurred, indicative of a drastic mitochondrial bottleneck, caused by massive colony mortality. In contrast, nuclear genetic diversity did not change, though hundreds of genes show signs of selection. The genetic diversity within each bee colony, particularly as a consequence of polyandry by queens, may enable preservation of genetic diversity even during population bottlenecks. These findings suggest that genetically diverse honey bee populations can recover from introduced diseases by evolving rapid tolerance, while maintaining much of the standing genetic variation.

Mating frequencies of honey bee queens (Apis mellifera L.) in a population of feral colonies in the Northeastern United States.

Across their introduced range in North America, populations of feral honey bee (Apis mellifera L.) colonies have supposedly declined in recent decades as a result of exotic parasites, most notably the ectoparasitic mite Varroa destructor. Nonetheless, recent studies have documented several wild populations of colonies that have persisted. The extreme polyandry of honey bee queens-and the increased intracolony genetic diversity it confers-has been attributed, in part, to improved disease resistance and may be a factor in the survival of these populations of feral colonies. We estimated the mating frequencies of queens in feral colonies in the Arnot Forest in New York State to determine if the level of polyandry of these queens is especially high and so might contribute to their survival success. We genotyped the worker offspring from 10 feral colonies in the Arnot Forest of upstate New York, as well as those from 20 managed colonies closest to this forest. We found no significant differences in mean mating frequency between the feral and managed queens, suggesting that queens in the remote, low-density population of colonies in the Arnot Forest are neither mate-limited nor adapted to mate at an especially high frequency. These findings support the hypothesis that the hyperpolyandry of honey bees has been shaped on an evolutionary timescale rather than on an ecological one.

Comparative brain transcriptomic analyses of scouting across distinct behavioural and ecological contexts in honeybees.

Individual differences in behaviour are often consistent across time and contexts, but it is not clear whether such consistency is reflected at the molecular level. We explored this issue by studying scouting in honeybees in two different behavioural and ecological contexts: finding new sources of floral food resources and finding a new nest site. Brain gene expression profiles in food-source and nest-site scouts showed a significant overlap, despite large expression differences associated with the two different contexts. Class prediction and 'leave-one-out' cross-validation analyses revealed that a bee's role as a scout in either context could be predicted with 92.5% success using 89 genes at minimum. We also found that genes related to four neurotransmitter systems were part of a shared brain molecular signature in both types of scouts, and the two types of scouts were more similar for genes related to glutamate and GABA than catecholamine or acetylcholine signalling. These results indicate that consistent behavioural tendencies across different ecological contexts involve a mixture of similarities and differences in brain gene expression.

A critical number of workers in a honeybee colony triggers investment in reproduction.

Social insect colonies, like individual organisms, must decide as they develop how to allocate optimally their resources among survival, growth, and reproduction. Only when colonies reach a certain state do they switch from investing purely in survival and growth to investing also in reproduction. But how do worker bees within a colony detect that their colony has reached the state where it is adaptive to begin investing in reproduction? Previous work has shown that larger honeybee colonies invest more in reproduction (i.e., the production of drones and queens), however, the term 'larger' encompasses multiple colony parameters including number of adult workers, size of the nest, amount of brood, and size of the honey stores. These colony parameters were independently increased in this study to test which one(s) would increase a colony's investment in reproduction via males. This was assayed by measuring the construction of drone comb, the special type of comb in which drones are reared. Only an increase in the number of workers stimulated construction of drone comb. Colonies with over 4,000 workers began building drone comb, independent of the other colony parameters. These results show that attaining a critical number of workers is the key parameter for honeybee colonies to start to shift resources towards reproduction. These findings are relevant to other social systems in which a group's members must adjust their behavior as a function of the group's size.

Local behavioral rules sustain the cell allocation pattern in the combs of honey bee colonies (Apis mellifera).

In the beeswax combs of honey bees, the cells of brood, pollen, and honey have a consistent spatial pattern that is sustained throughout the life of a colony. This spatial pattern is believed to emerge from simple behavioral rules that specify how the queen moves, where foragers deposit honey/pollen and how honey/pollen is consumed from cells. Prior work has shown that a set of such rules can explain the formation of the allocation pattern starting from an empty comb. We show that these rules cannot maintain the pattern once the brood start to vacate their cells, and we propose new, biologically realistic rules that better sustain the observed allocation pattern. We analyze the three resulting models by performing hundreds of simulation runs over many gestational periods and a wide range of parameter values. We develop new metrics for pattern assessment and employ them in analyzing pattern retention over each simulation run. Applied to our simulation results, these metrics show alteration of an accepted model for honey/pollen consumption based on local information can stabilize the cell allocation pattern over time. We also show that adding global information, by biasing the queen's movements towards the center of the comb, expands the parameter regime over which pattern retention occurs.

No facultative worker policing in the honey bee (Apis mellifera L.).

Kin selection theory predicts that in colonies of social Hymenoptera with multiply mated queens, workers should mutually inhibit ("police") worker reproduction, but that in colonies with singly mated queens, workers should favor rearing workers' sons instead of queens' sons. In line with these predictions, Mattila et al. (Curr Biol 22:2027-2031, 2012) documented increased ovary development among workers in colonies of honey bees with singly mated queens, suggesting that workers can detect and respond adaptively to queen mating frequency and raising the possibility that they facultative police. In a follow-up experiment, we test and reject the hypothesis that workers in single-patriline colonies prefer worker-derived males and are able to reproduce directly; we show that their eggs are policed as strongly as those of workers in colonies with multiply mated queens. Evidently, workers do not respond facultatively to a kin structure that favors relaxed policing and increased direct reproduction. These workers may instead be responding to a poor queen or preparing for possible queen loss.

Beeping and piping: characterization of two mechano-acoustic signals used by honey bees in swarming.

Of the many signals used by honey bees during the process of swarming, two of them--the stop signal and the worker piping signal--are not easily distinguished for both are mechano-acoustic signals produced by scout bees who press their bodies against other bees while vibrating their wing muscles. To clarify the acoustic differences between these two signals, we recorded both signals from the same swarm and at the same time, and compared them in terms of signal duration, fundamental frequency, and frequency modulation. Stop signals and worker piping signals differ in all three variables: duration, 174 ± 64 vs. 602 ± 377 ms; fundamental frequency, 407 vs. 451 Hz; and frequency modulation, absent vs. present. While it remains unclear which differences the bees use to distinguish the two signals, it is clear that they do so for the signals have opposite effects. Stop signals cause inhibition of actively dancing scout bees whereas piping signals cause excitation of quietly resting non-scout bees.

Molecular determinants of scouting behavior in honey bees.

Little is known about the molecular basis of differences in behavior among individuals. Here we report consistent novelty-seeking behavior, across different contexts, among honey bees in their tendency to scout for food sources and nest sites, and we reveal some of the molecular underpinnings of this behavior relative to foragers that do not scout. Food scouts showed extensive differences in brain gene expression relative to other foragers, including differences related to catecholamine, glutamate, and γ-aminobutyric acid signaling. Octopamine and glutamate treatments increased the likelihood of scouting, whereas dopamine antagonist treatment decreased it. These findings demonstrate intriguing similarities in human and insect novelty seeking and suggest that this trait, which presumably evolved independently in these two lineages, may be subserved by conserved molecular components.

Stop signals provide cross inhibition in collective decision-making by honeybee swarms.

Honeybee swarms and complex brains show many parallels in how they make decisions. In both, separate populations of units (bees or neurons) integrate noisy evidence for alternatives, and, when one population exceeds a threshold, the alternative it represents is chosen. We show that a key feature of a brain--cross inhibition between the evidence-accumulating populations--also exists in a swarm as it chooses its nesting site. Nest-site scouts send inhibitory stop signals to other scouts producing waggle dances, causing them to cease dancing, and each scout targets scouts' reporting sites other than her own. An analytic model shows that cross inhibition between populations of scout bees increases the reliability of swarm decision-making by solving the problem of deadlock over equal sites.

Kin selection and eusociality.

Arising from M. A. Nowak, C. E. Tarnita & E. O. Wilson 466, 1057-1062 (2010); Nowak et al. reply. Hamilton described a selective process in which individuals affect kin (kin selection), developed a novel modelling strategy for it (inclusive fitness), and derived a rule to describe it (Hamilton's rule). Nowak et al. assert that inclusive fitness is not the best modelling strategy, and also that its production has been "meagre". The former may be debated by theoreticians, but the latter is simply incorrect. There is abundant evidence to demonstrate that inclusive fitness, kin selection and Hamilton's rule have been extraordinarily productive for understanding the evolution of sociality.

Sleep deprivation impairs precision of waggle dance signaling in honey bees.

Sleep is essential for basic survival, and insufficient sleep leads to a variety of dysfunctions. In humans, one of the most profound consequences of sleep deprivation is imprecise or irrational communication, demonstrated by degradation in signaling as well as in receiving information. Communication in nonhuman animals may suffer analogous degradation of precision, perhaps with especially damaging consequences for social animals. However, society-specific consequences of sleep loss have rarely been explored, and no function of sleep has been ascribed to a truly social (eusocial) organism in the context of its society. Here we show that sleep-deprived honey bees (Apis mellifera) exhibit reduced precision when signaling direction information to food sources in their waggle dances. The deterioration of the honey bee's ability to communicate is expected to reduce the foraging efficiency of nestmates. This study demonstrates the impact of sleep deprivation on signaling in a eusocial animal. If the deterioration of signals made by sleep-deprived honey bees and humans is generalizable, then imprecise communication may be one detrimental effect of sleep loss shared by social organisms.

No intracolonial nepotism during colony fissioning in honey bees.

Most species of social insects have singly mated queens, but in some species each queen mates with numerous males to create a colony whose workers belong to multiple patrilines. This colony genetic structure creates a potential for intracolonial nepotism. One context with great potential for such nepotism arises in species, like honey bees, whose colonies reproduce by fissioning. During fissioning, workers might nepotistically choose between serving a young (sister) queen or the old (mother) queen, preferring the former if she is a full-sister but the latter if the young queen is only a half-sister. We examined three honeybee colonies that swarmed, and performed paternity analyses on the young (immature) queens and samples of workers who either stayed with the young queens in the nest or left with the mother queen in the swarm. For each colony, we checked whether patrilines represented by immature queens had higher proportions of staying workers than patrilines not represented by immature queens. We found no evidence of this. The absence of intracolonial nepotism during colony fissioning could be because the workers cannot discriminate between full-sister and half-sister queens when they are immature, or because the costs of behaving nepotistically outweigh the benefits.

Independence and interdependence in collective decision making: an agent-based model of nest-site choice by honeybee swarms.

Condorcet's jury theorem shows that when the members of a group have noisy but independent information about what is best for the group as a whole, majority decisions tend to outperform dictatorial ones. When voting is supplemented by communication, however, the resulting interdependencies between decision makers can strengthen or undermine this effect: they can facilitate information pooling, but also amplify errors. We consider an intriguing non-human case of independent information pooling combined with communication: the case of nest-site choice by honeybee (Apis mellifera) swarms. It is empirically well documented that when there are different nest sites that vary in quality, the bees usually choose the best one. We develop a new agent-based model of the bees' decision process and show that its remarkable reliability stems from a particular interplay of independence and interdependence between the bees.

Sensory coding of nest-site value in honeybee swarms.

This study investigates the first stage of the decision-making process of a honeybee swarm as it chooses a nest site: how a scout bee codes the value of a potential nest site in the waggle dances she produces to represent this site. We presented honeybee swarms with a two-alternative choice between a high-value site and a medium-value site and recorded the behavior of individually identifiable scout bees as they reported on these two alternatives. We found that bees performed equally lengthy inspections at the two sites, but that, on the swarm cluster, they performed more dance circuits per bee for the high-value site. We also found that there was much individual-level noise in the coding of site value, but that there were clear population-level differences in total dance circuits produced for the two sites. The first bee to find a site had a high probability of reporting the site with a waggle dance, regardless of its value. This discoverer-should-dance phenomenon may help ensure that a swarm gives attention to all discovered sites. There was rapid decay in the dance response; the number of dance circuits produced by a bee after visiting a site decreased linearly over sequential visits, and eventually each bee ceased visiting her site. This decay, or ;leakage', in the accumulation of bees at a site improves a swarm's decision-making ability by helping a swarm avoid making fast-decision errors.