Honeybee Colony Thermoregulation – Regulatory Mechanisms and Contribution of Individuals in Dependence on Age,Location and Thermal Stress

Honeybee larvae and pupae are extremely stenothermic, i.e. they strongly depend on accurate regulation of brood nest temperature for proper development (33–36°C). Here we study the mechanisms of social thermoregulation of honeybee colonies under changing environmental temperatures concerning the contribution of individuals to colony temperature homeostasis. Beside migration activity within the nest, the main active process is “endothermy on demand” of adults. An increase of cold stress (cooling of the colony) increases the intensity of heat production with thoracic flight muscles and the number of endothermic individuals, especially in the brood nest. As endothermy means hard work for bees, this eases much burden of nestmates which can stay ectothermic. Concerning the active reaction to cold stress by endothermy, age polyethism is reduced to only two physiologically predetermined task divisions, 0 to ∼2 days and older. Endothermic heat production is the job of bees older than about two days. They are all similarly engaged in active heat production both in intensity and frequency. Their active heat production has an important reinforcement effect on passive heat production of the many ectothermic bees and of the brood. Ectothermy is most frequent in young bees (<∼2 days) both outside and inside of brood nest cells. We suggest young bees visit warm brood nest cells not only to clean them but also to speed up flight muscle development for proper endothermy and foraging later in their life. Young bees inside brood nest cells mostly receive heat from the surrounding cell wall during cold stress, whereas older bees predominantly transfer heat from the thorax to the cell wall. Endothermic bees regulate brood comb temperature more accurately than local air temperature. They apply the heat as close to the brood as possible: workers heating cells from within have a higher probability of endothermy than those on the comb surface. The findings show that thermal homeostasis of honeybee colonies is achieved by a combination of active and passive processes. The differential individual endothermic and behavioral reactions sum up to an integrated action of the honeybee colony as a superorganism.     

Dynamics of Collective Decision Making of Honeybees in Complex Temperature Fields

Endothermic heat production is a crucial evolutionary adaptation that is, amongst others, responsible for the great success of honeybees. Endothermy ensures the survival of the colonies in harsh environments and is involved in the maintenance of the brood nest temperature, which is fundamental for the breeding and further development of healthy individuals and thus the foraging and reproduction success of this species. Freshly emerged honeybees are not yet able to produce heat endothermically and thus developed behavioural patterns that result in the location of these young bees within the warm brood nest where they further develop and perform tasks for the colony. Previous studies showed that groups of young ectothermic honeybees exposed to a temperature gradient collectively aggregate at the optimal place with their preferred temperature of 36°C but most single bees do not locate themselves at the optimum. In this work we further investigate the behavioural patterns that lead to this collective thermotaxis. We tested single and groups of young bees concerning their ability to discriminate a local from a global temperature optimum and, for groups of bees, analysed the speed of the decision making process as well as density dependent effects by varying group sizes. We found that the majority of tested single bees do not locate themselves at the optimum whereas sufficiently large groups of bees are able to collectively discriminate a suboptimal temperature spot and aggregate at 36°C. Larger groups decide faster than smaller ones, but in larger groups a higher percentage of bees may switch to the sub-optimum due to crowding effects. We show that the collective thermotaxis is a simple but well evolved, scalable and robust social behaviour that enables the collective of bees to perform complex tasks despite the limited abilities of each individual.     

Nonrandom visitation of brood cells by worker honey bees (Hymenoptera: Apidae)

The visitation pattern by worker honey bees to cells in the brood nest was monitored on an artificially created brood pattern consisting of about one-fourth brood cells evenly distributed among empty cells. The majority (63 %) of the observed workers selectively entered larval cells. In contrast, some workers avoided egg cells when presented a choice of egg vs empty cells. The results suggest that larvae produce a general signal indicating their presence to worker bees. Eggs also seem to produce a signal, which is perceived to be different from the one from larvae.     

Pupal developmental temperature and behavioral specialization of honeybee workers (<Emphasis Type="Italic">Apis mellifera</Emphasis> L.)

Honeybees (Apis mellifera) are able to regulate the brood nest temperatures within a narrow range between 32 and 36°C. Yet this small variation in brood temperature is sufficient to cause significant differences in the behavior of adult bees. To study the consequences of variation in pupal developmental temperature we raised honeybee brood under controlled temperature conditions (32, 34.5, 36°C) and individually marked more than 4,400 bees, after emergence. We analyzed dancing, undertaking behavior, the age of first foraging flight, and forager task specialization of these workers. Animals raised under higher temperatures showed an increased probability to dance, foraged earlier in life, and were more often engaged in undertaking. Since the temperature profile in the brood nest may be an emergent property of the whole colony, we discuss how pupal developmental temperature can affect the overall organization of division of labor among the individuals in a self-organized process.     

Short-term fumigation of honey bee (Hymenoptera: Apidae) colonies with formic and acetic acids for the control of Varroa destructor (Acari: Varroidae)

Controlling populations of varroa mites is crucial for the survival of the beekeeping industry. Many treatments exist, and all are designed to kill mites on adult bees. Because the majority of mites are found under capped brood, most treatments are designed to deliver active ingredients over an extended period to control mites on adult bees, as developing bees and mites emerge. In this study, a 17-h application of 50% formic acid effectively killed mites in capped worker brood and on adult bees without harming queens or uncapped brood. Neither acetic acid nor a combined treatment of formic and acetic acids applied to the West Virginia formic acid fumigator was as effective as formic acid alone in controlling varroa mites. In addition, none of the treatments tested in late summer had an effect on the late-season prevalence of deformed wing virus. The short-term formic acid treatment killed > 60% of varroa mites in capped worker brood; thus, it is a promising tool for beekeepers, especially when such treatments are necessary during the nectar flow.     

Nestmate recognition in the stingless bee Melipona panamica (Apidae, Meliponini)

Summary: Nestmate recognition was studied in the Neotropical stingless bee Melipona panamica, a species in which workers "sneak" their own reproductive eggs into 1 % of brood cells. We manipulated four factors that could influence individual recognition cues: the mother queen, the environment during the immature stage, the environment during the early adult stage, and worker age. We also simulated the action of natural enemies on colonies tested for discrimination of such worker characteristics. All factors that we tested affected responses of the discriminating workers, which could recognize sisters, nieces and unrelated workers. Previous exposure of unrelated callow bees to the odor of the host nest greatly increased chances of acceptance by the host colony. Probability of acceptance decreased, however, with increasing age of introduced bees or increasing disturbance of the host colony. These complexities in patterns of nestmate recognition and nest defense are adequately explained from the standpoint of inclusive fitness of the discriminating workers. Differences in nestmate recognition and worker egg laying among Meliponini are also discussed.     

Varroa destructor infestation in untreated honey bee (Hymenoptera: Apidae) colonies selected for hygienic behavior

Honey bee (Apis mellifera L.) colonies bred for hygienic behavior were tested in a large field trial to determine if they were able to resist the parasitic mite Varroa destructor better than unselected colonies of"Starline" stock. Colonies bred for hygienic behavior are able to detect, uncap, and remove experimentally infested brood from the nest, although the extent to which the behavior actually reduces the overall mite-load in untreated, naturally infested colonies needed further verification. The results indicate that hygienic colonies with queens mated naturally to unselected drones had significantly fewer mites on adult bees and within worker brood cells than Starline colonies for up to 1 yr without treatment in a commercial, migratory beekeeping operation. Hygienic colonies actively defended themselves against the mites when mite levels were relatively low. At high mite infestations (>15% of worker brood and of adult bees), the majority of hygienic colonies required treatment to prevent collapse. Overall, the hygienic colonies had similar adult populations and brood areas, produced as much honey, and had less brood disease than the Starline colonies. Thus, honey bees bred for hygienic behavior performed as well if not better than other commercial lines of bees and maintained lower mite loads for up to one year without treatment.     

Comparative analysis of thermoregulation in the nests of certain species of social insects

Studies have been made on thermal regulation in the nests of families of the honey bee Apis mellifera, wasp Dolihovespula silvestris and bumblebees Bombus terrestris, B. agrorum and B. lapidaris during their maximum development. It was shown that thermoregulation significantly stimulated the brood in the nest. Among the species investigated, the highest thermoregulatory capacities are exhibited by honey bees, this fact being associated with the large number of individuals in their colonies. During cooling, bees group around the brood and their bodies make a cover of thermal insulation. Sheltering of the nests plays the main role in heat preservation of wasps and bumblebees. The latter, using contact method of heating of cells with the brood, provide rather constant temperature for its development. Honey bees, wasps and bumblebees react to overheating in the nests essentially in a similar way, i. e. by active aeration of their nests by vigorous wing beatings. The frequency of beatings increases with the increase in temperature.     

Effects of Brood Pheromone Modulated Brood Rearing Behaviors on Honey Bee (Apis mellifera L.) Colony Growth

A hallmark of eusociality is cooperative brood care. In most social insect systems brood rearing labor is divided between individuals working in the nest tending the queen and larvae, and foragers collecting food outside the nest. To place brood rearing division of labor within an evolutionary context, it is necessary to understand relationships between individuals in the nest engaged in brood care and colony growth in the honey bee. Here we examined responses of the queen, queen-worker interactions, and nursing behaviors to an increase in the brood rearing stimulus environment using brood pheromone. Colony pairs were derived from a single source and were headed by open-mated sister queens, for a total of four colony pairs. One colony of a pair was treated with 336 μg of brood pheromone, and the other a blank control. Queens in the brood pheromone treated colonies laid significantly more eggs, were fed longer, and were less idle compared to controls. Workers spent significantly more time cleaning cells in pheromone treatments. Increasing the brood rearing stimulus environment with the addition of brood pheromone significantly increased the tempo of brood rearing behaviors by bees working in the nest resulting in a significantly greater amount of brood reared.     

Modelling the role of intracolonial genetic diversity on regulation of brood temperature in honey bee (Apis mellifera L.) colonies

In polyandrous social insects such as honey bees, a worker’s affinity for a particular task may be genetically infl uenced and so some patrilines may have lower stimulus thresholds for commencing a task than others. We used simulation models to investigate the effects of intracolonial diversity in the task thresholds that stimulate workers to engage in heating and cooling during nest thermoregulation. First, we simulated colonies comprised of one or 15 patrilines that were engaged in heating the brood nest, and observed that single patriline colonies maintained, on average, less stable brood nest temperatures than multiple patriline colonies. Second we simulated colonies with five patrilines that were engaged in cooling their nest, recording the proportions of bees of different patrilines that engaged in nest cooling in response to changing temperatures. Both of our simulations show remarkably similar qualitative patterns to those that we have previously observed empirically. This provides further support for the hypothesis that geneticallybased variability in task thresholds among patrilines within honey bee colonies is an important contributor to the ability of colonies to precisely thermoregulate their nests, and we suggest that diversity is important for optimal expression of a range of other colony-level phenotypes. Received 17 June 2005; revised 27 October 2005; accepted 23 December 2005.     

It's good to be queen: classically eusocial colony structure and low worker fitness in an obligately social sweat bee

Lasioglossum malachurum, a bee species common across much of Europe, is obligately eusocial across its range but exhibits clear geographic variation in demography and social behaviour. This variation suggests that social interactions between queens and workers, opportunities for worker oviposition, and patterns of relatedness among nest mates may vary considerably, both within and among regions. In this study, we used three microsatellite loci with 12-18 alleles each to examine the sociogenetic structure of colonies from a population at Agios Nikolaos Monemvasias in southern Greece. These analyses reveal that the majority of colonies exhibit classical eusocial colony structure in which a single queen mated to a single male monopolizes oviposition. Nevertheless, we also detect low rates of multiqueen nest founding, occasional caste switching by worker-destined females, and worker oviposition of both gyne and male-producing eggs in the final brood. Previous evidence that the majority of workers show some ovarian development and a minority (17%) have at least one large oocyte contrasts with the observation that only 2-3% of gynes and males (the so-called reproductive brood) are produced by workers. An evaluation of the parameters of Hamilton's Rule suggests that queens benefit greatly from the help provided by workers but that workers achieve greater fitness by provisioning and laying their own eggs rather than by tending to the queen's eggs. This conflict of interest between the queen and her workers suggests that the discrepancy between potential and achieved worker oviposition is due to queen interference. Comparison of relatedness and maternity patterns in the Agios Nikolaos Monemvasias population with those from a northern population near Tübingen, Germany, points to a north-south cline of increasingly effective queen control of worker behaviour.     

Calorimetric investigations of the different castes of honey bees,Apis mellifera carnica

Summary Honey bees of different age and castes were investigated calorimetrically at 20, 25 and 30 °C. Experiments were completed by endoscopic observation of the insects in the visible and the near infrared range and by acoustical monitoring and subsequent frequency analysis of various locomotor activities. Direct calorimetric results of this paper are compared with data of indirect calorimetry from the literature using a respiratory quotient of 1.00 and 21.13 J consumed. Agreements between both methods are generally good. The results show that weight-specific heat production rates increase with age of worker bees by a factor of 5.6 at 30 °C, 3.7 at 25 °C and 40.0 at 20 °C. In groups of foragers the heat production decreases with growing group size to around 6% of the value for an isolated bee. The presence of a fertile queen or of brood reduces the heat output of a small worker group significantly. Adult drones exhibit a much higher metabolic rate (up to 19.7-fold at 20 °C) than juveniles with strong fluctuations in the power-time curves. Fertile queens show a less pronounced heat production rate than virgin queens (54% at 30 °C, 87% at 25 °C and 77% at 20 °C). Calorimetric unrest is much higher for young than for adult queens. Heat production is very low in both uncapped and capped brood and less than 30% of that of a newly emerged worker. In most cases temperature showed a significant influence on the metabolic level, although its sign was not homogeneous between the castes or even within them. Locomotor activities are easily recorded by the acoustic frequency spectrum (0–7.5 kHz) and in good agreement with endoscopic observations and calorimetric traces.Abbreviations RQ respiratory quotient - ww wet weight This paper is part of the PhD thesis of L.F.     

Thresholds of Response in Nest Thermoregulation by Worker Bumble Bees, Bombus bifarius nearcticus (Hymenoptera: Apidae)

Regulation of nest temperature is important to the fitness of eusocial insect colonies. To maintain appropriate conditions for the developing brood, workers must exhibit thermoregulatory responses to ambient temperature. Because nest-mate workers differ in task performance, thermoregulatory behavior provides an opportunity to test threshold of response models for the regulation of division of labor. We found that worker bumble bees ( Bombus bifarius nearcticus ) responded to changes in ambient temperature by altering their rates of performing two tasks – wing fanning and brood cell incubation. At the colony level, the rate of incubating decreased, and the rate of fanning increased, with increasing temperature. Changes in the number of workers performing these tasks were more important to the colony response than changes in workers' task performance rates. At the individual level, workers' lifetime rates of incubation and fanning were positively correlated, and most individuals did not specialize exclusively on either of these temperature-sensitive tasks. However, workers differed in the maximum temperature at which they incubated and in the minimum temperature at which they fanned. More individuals fanned at high and incubated at low temperatures. Most of the workers that began incubating at higher temperatures continued performing this task at lower temperatures, when additional nest-mates became active. The converse was true for fanning behavior. These data are consistent with a threshold of response model for thermoregulatory behavior of B. bifarius workers.     

Sociometry and sociogenesis of colonies of the harvester ant, Pogonomyrmex badius: distribution of workers, brood and seeds within the nest in relation to colony size and season

1. The vertical distribution in the nest of chambers, workers, callow workers, brood and seeds was studied in the harvester ant, Pogonomyrmex badius, in northern Florida. On each of four sample dates (May, July, October, January), six to seven colonies, chosen to represent the full range of sizes, were excavated. All chamber contents were collected and counted. Chambers were mapped and measured. In a preliminary study, two nests were excavated after preventing vertical migration by driving barriers into the wall of a pit next to the nest, severing the vertical tunnels. The vertical distribution of these barrier-nests differed little from unrestrained nests, indicating that unrestrained excavation produced a reasonable picture of vertical distributions. 2. Nest depth, chamber number and total area increased with colony size. Chamber area declined sharply with depth, as did chamber number, such that more than half of the total area was found in the upper quarter of the nest. 3. The proportion of dark-coloured (older) workers also declined strongly with depth, but this decline was weaker in the spring, and depended to a modest degree on colony size. Conversely, in the distribution of callow (young) workers, the proportion increased towards the bottom of the nest. Mean worker age was inversely related to the depth at which workers were found. The proportion of the brood also increased towards the bottom of the nest, with worker brood, sexual brood, pupae and larvae all being distributed similarly. 4. By contrast, seeds were stored at a preferred absolute depth between 40 and 100 cm. Colonies shallower than 100 cm stored seeds in their deepest chambers. Larger colonies stored most seeds in the upper third of the nest, but patterns were somewhat erratic because chambers were either filled completely with seeds or were empty. 5. Because chamber area decreased sharply with depth, the densities (individuals cm^–2) of all colony members, including dark workers, were lowest near the surface and highest in the deepest parts of the nest. Here, worker densities ranged from 2 to 8 cm^–2, and brood from 2 to 25 cm^–2. 6. The regularity of the patterns of distribution suggests that harvester ant colonies have considerable spatial and temporal structure, which serves or is the outcome of important colony processes. A simple mechanism that could generate several of these patterns is discussed. New workers produced deep in the nest move upwards as they age. As they leave the brood zone they change from brood care to general nest duties, including increased nest excavation, leading to the top-heavy pattern of nest area. As they appear at the surface, they change to guarding and foraging. Thus, age polyethism may be partly the result of this upward migration of workers.     

Some observations on the concurrent parasitization ofApis florea byTropilaelaps clareae andEuvarroa sinhai

Adult bees, worker brood cells and drone brood cells ofApis florea were examined for the presence of mites by stereo microscope and by washing with soap. Tropilaelaps clareae was only found on adult bees;Euvarroa sinhai on adult bees and drone brood. The level ofT. clareae infestation is always very low, generally not exceeding 0.1%; that ofE. sinhai is somewhat higher. The mites were never found together on a single bee.     

Kin recognition of worker brood by worker honey bees,Apis mellifera L.

Experimental colonies of honey bees consisting of two patrilines were observed as they reared worker brood. Seven behavior patterns that relate to brood care were recorded. Worker bees biased the care they provided to eggs and larvae destined to become workers on the basis of brood patrilines. Both patrilineal and antipatrilineal preferences in various behavioral patterns were observed. There was variation among colonies that may have been the result of the frequencies of brood of each patriline and the total amount of brood available to be reared. In addition, there were some differences between workers of the two patrilines in the way that they cared for the two patrilines of brood.     

Ergonomics in stingless bees: changes in intranidal behavior after partial removal of storage pots and honey inMelipona favosa (Hym. Apidae,Meliponinae)

Summary The ergonomics of task allocation in stingless bees was examined in two laboratory colonies ofMelipona favosa containing individually marked workers. Performance of intranidal behavior patterns, brood cell production and the processing and storage of nectar, were studied during a control and a wax-deprived period. Experimental wax deprivation consisted of regular controlled removal of storage pots and their contents. Stored carbohydrate concentration and storage pot mass declined during the wax-deprived period. Behavior was measured by sampling for a complete ethogram at seven intranidal areas.In comparison withApis mellifera, M. favosa appeared to be more sensitive to stress, with a distinct reallocation of effort between tasks. The significant increase under stress of brood production and brood-rearing behavior patterns observed was previously also found inApis. Self-oriented behavior patterns declined under stressed circumstances.Involucrum construction declined in wax-deprivedMelipona colonies. Relatively low rates of walking behavior in general inMelipona suggest a low mean free distance between tasks. These results are of particular interest in relation to the basically different nest structure of stingless bees, e.g. very distinct areas for brood production and the processing and storage of food. This architecture of stingless bee nests appeared to influence strongly their specific responses to wax deprivation.     

Major benefits of guarding behavior in subsocial bees: implications for social evolution

Parental care is a behavior that increases the growth and survival of offspring, often at a cost to the parents' own survival and/or future reproduction. In this study, we focused on nest guarding, which is one of the most important types of extended parental care; we studied this behavior in two solitary bee species of the genus Ceratina with social ancestors. We performed the experiment of removing the laying female, who usually guards the nest after completing its provisioning, to test the effects of nest guarding on the offspring survival and nest fate. By dissecting natural nests, we found that Ceratina cucurbitina females always guarded their offspring until the offspring reached adulthood. In addition, the females of this species were able to crawl across the nest partitions and inspect the offspring in the brood cells. In contrast, several Ceratina chalybea females guarded their nests until the offspring reached adulthood, but others closed the nest entrance with a plug and deserted the nest. Nests with a low number of provisioned cells were more likely to be plugged and abandoned than nests with a higher number of cells. The female removal experiment had a significantly negative effect on offspring survival in both species. These nests frequently failed due to the attacks of natural enemies (e.g., ants, chalcidoid wasps, and other competing Ceratina bees). Increased offspring survival is the most important benefit of the guarding strategy. The abandonment of a potentially unsuccessful brood might constitute a benefit of the nest plugging behavior. The facultative nest desertion strategy is a derived behavior in the studied bees and constitutes an example of an evolutionary reduction in the extent of parental care.     

ORIGINS,EVOLUTION, AND DIVERSIFICATION OF CLEPTOPARASITIC LINEAGES IN LONG‐TONGUED BEES

The evolution of parasitic behavior may catalyze the exploitation of new ecological niches yet also binds the fate of a parasite to that of its host. It is thus not clear whether evolutionary transitions from free‐living organism to parasite lead to increased or decreased rates of diversification. We explore the evolution of brood parasitism in long‐tongued bees and find decreased rates of diversification in eight of 10 brood parasitic clades. We propose a pathway for the evolution of brood parasitic strategy and find that a strategy in which a closed host nest cell is parasitized and the host offspring is killed by the adult parasite represents an obligate first step in the appearance of a brood parasitic lineage; this ultimately evolves into a strategy in which an open host cell is parasitized and the host offspring is killed by a specialized larval instar. The transition to parasitizing open nest cells expanded the range of potential hosts for brood parasitic bees and played a fundamental role in the patterns of diversification seen in brood parasitic clades. We address the prevalence of brood parasitic lineages in certain families of bees and examine the evolution of brood parasitism in other groups of organisms.     

A test of adaptive hypotheses for rapid nest construction in a swarm-founding wasp

Most social insect species enlarge their nests gradually and in close correlation with the growing need for space for brood and/or stored food. In contrast, some species of swarm-founding eusocial wasps construct the nest rapidly to a final size in the first two to three weeks of the founding stage. We considered four hypotheses on the functions of rapid nest construction in the wasp Polybia occidentalis and directly tested two of them. The first hypothesis is that rapid construction maximizes output of the worker force when there are few other work demands; it predicts that construction rate remains high until the first eggs begin to hatch, following which it declines as increasing amounts of worker effort are allocated to the feeding of larvae. The second says that rapid nest construction minimizes the time the adults in the swarm are exposed to predation and the elements; it predicts that nest-construction rate should drop steeply after the nest is large enough to house all the adults in the swarm. We measured pulp-foraging rates for the first 12 days of the founding stage in control colonies and in colonies whose nests we manipulated to prevent housing of the swarm. The treatment and control groups did not differ in construction rate for several days following the housing event, contradicting the adult-protection hypothesis. Late in nest construction, treatment colonies were building at significantly higher rates than were control colonies. If demand for brood care were a major factor in determining construction rate, both groups would have responded to the eclosion of larvae in the same way and shown a parallel decline in construction rate, but this did not happen. Instead, the patterns of nest construction rate we observed provided indirect support for the two remaining hypotheses. The first of these is that rapid construction minimizes exposure of the brood to natural enemies and desiccation. The second is that rapid construction promotes competition among queens by providing empty cells for oviposition, thereby facilitating the selecting out of the less fecund of the multiple reproductive females. Also consistent with this hypothesis is the apparent absence of explosive nest construction in monogynous, eusocial bees. Received 13 October 2007; revised 31 March 2008; accepted 6 April 2008.