No evidence that reproductive bumblebee workers reduce the production of new queens

Kin selection theory predicts potential conflict between queen and workers over male parentage in hymenopteran societies headed by one, singly mated queen, because each party is more closely related to its own male offspring. In ‘late-switching’ colonies of the bumblebee Bombus terrestris, i.e. colonies whose queens lay haploid eggs relatively late in the colony cycle, workers start to lay male eggs shortly after the queen lays the female eggs that will develop into new queens. It has been hypothesized that this occurs because workers recognize, via a signal given by the queen instructing female larvae to commence development as queens, that egg laying is now in their kin-selected interest. This hypothesis assumes that aggressive behaviour in egg-laying workers does not substantially reduce the production of new queens, which would decrease the workers' fitness payoff from producing males. We tested the hypothesis that reproductive activity inB. terrestris workers does not reduce the production of new queens. We used microsatellite genotyping to sex eggs and hence to select eight size-matched pairs of ‘late-switching’ colonies from a set of commercial colonies. From one colony of each pair we removed every egg-laying or aggressive worker observed. From the other colony, we simultaneously removed a nonegg-laying, nonaggressive worker. Removed workers were replaced with young workers from separate colonies at equal frequencies within the pair. There was no significant difference in queen productivity between colonies with reduced or normal levels of egg-laying or aggressive workers. Therefore, as predicted, reproductive B. terrestris workers did not significantly reduce the production of new queens.     

Queen execution in a monogynous ant

Workers in many species of social insects are capable of laying unfertilized eggs, which can develop into haploid males. This causes a conflict about male parentage between queens and workers. In a few species, this may result in matricide, that is, workers kill the colony''s queen. Queen killing has so far been observed mainly in multi‐queen colonies or in annual species, when the queen''s fecundity declines at the end of the reproductive period. Here, we report queen expulsion and matricide in a monogynous, monandrous ant with perennial societies. Workers were seen to aggressively expel both related and unrelated queens from their nest shortly after the end of hibernation. Queen expulsion and matricide led to a significant decrease in the number of workers and brood, but eventually increased the direct fitness of workers through significant male production. Long‐term observations revealed a short lifespan of queens, while workers in orphaned colonies survived and produced male offspring over several years.     

QUEEN NUMBER IN COLONIES OF SOCIAL HYMENOPTERA AS A KIN-SELECTED ADAPTATION

Using a series of kin-selection models, I examine factors that favor multiple egg-laying queens (polygyny) in eusocial Hymenoptera colonies. One result is that there is a theoretical conflict of interest between the founding queens and their daughter workers over how many and which individuals should be the extra reproductives. Both castes should prefer their full sisters. Therefore, primary polygyny (multiple related foundresses) may favor queens while secondary polygyny (related queens added to mature colonies) may favor workers. Polygyny, itself, was found to be favored by high colony survivorship and low probability of queens contributing eggs to successive broods. Polygyne colonies, however, did not need to produce more offspring per brood to be selectively favored; they could be half as productive per brood as monogyne ones and still have higher lifetime fitness under some conditions. For reproductive data from eight ant species with both monogyne and polygyne colonies, the model generates results that are consistent with a kin-selection explanation of polygyny in all of them. It is proposed that queen number is an ecologically flexible trait that is influenced by a broad set of factors but is not necessarily linked to specific habitat types. Furthermore, neither polygyny nor monogyny may be reliably considered as the primitive or ancestral Hymenopteran social system. The optimal queen number within a species may evolutionarily increase or decrease, depending on the direction of environmental change.     

Clustered male production by workers in the stingless bee Melipona subnitida Ducke (Apidae, Meliponinae)

Summary: In stingless bees brood cells are sequentially filled with liquid larval food (mass-provisioning), upon which the queen lays an egg. Thereafter the cell is closed by a worker. This study showed that during these processes workers of Melipona subnitida regularly laid eggs that served as food for the queen. Occasionally cells were oviposited in and immediately closed by a worker. These cells always rendered males. Some of these reproductive workers were seen to lay a trophic egg as well. Cells which were exclusively oviposited in by the physogastric queen gave rise to workers and queens only. In one colony it could be verified that three workers alone, which differed in age by one day, laid 15 male-producing eggs within a period of two successive weeks. Among them the number of ovipositions was positively related to the order in which workers eclosed - the oldest worker laying most eggs - and inversely related to the number of times they closed cells oviposited in exclusively by the queen. Apparently the physogastric queen was not able to stop certain workers from reproducing. We therefore conclude that some workers in M. subnitida temporarily dominated their queens in egg-laying.     

Worker reproduction and policing in insect societies: an ESS analysis

Insect societies are vulnerable to exploitation by workers who reproduce selfishly rather than help to rear the queen's offspring. In most species, however, only a small proportion of the workers reproduce. Here, we develop an evolutionarily stable strategy (ESS) model to investigate factors that could explain these observed low levels of reproductive exploitation. Two key factors are identified: relatedness and policing. Relatedness affects the ESS proportion of reproductive workers because laying workers generally work less, leading to greater inclusive fitness costs when within-colony relatedness is higher. The second key factor is policing. In many species, worker-laid eggs are selectively removed or 'policed' by other workers or the queen. We show that policing not only prevents the rearing of worker-laid eggs but can also make it unprofitable for workers to lay eggs in the first place. This can explain why almost no workers reproduce in species with efficient policing, such as honeybees, Apis, and the common wasp, Vespula vulgaris, despite relatively low relatedness caused by multiple mating of the mother queen. Although our analyses focus on social insects, the conclusion that both relatedness and policing can reduce the incentive for cheating applies to other biological systems as well.     

Social Context and Reproductive Potential Affect Worker Reproductive Decisions in a Eusocial Insect

Context-dependent decision-making conditions individual plasticity and is an integrant part of alternative reproductive strategies. In eusocial Hymenoptera (ants, bees and wasps), the discovery of worker reproductive parasitism recently challenged the view of workers as a homogeneous collective entity and stressed the need to consider them as autonomous units capable of elaborate choices which influence their fitness returns. The reproductive decisions of individual workers thus need to be investigated and taken into account to understand the regulation of reproduction in insect societies. However, we know virtually nothing about the proximate mechanisms at the basis of worker reproductive decisions. Here, we test the hypothesis that the capacity of workers to reproduce in foreign colonies lies in their ability to react differently according to the colonial context and whether this reaction is influenced by a particular internal state. Using the bumble bee Bombus terrestris, we show that workers exhibit an extremely high reproductive plasticity which is conditioned by the social context they experience. Fertile workers reintroduced into their mother colony reverted to sterility, as expected. On the contrary, a high level of ovary activity persisted in fertile workers introduced into a foreign nest, and this despite more frequent direct contacts with the queen and the brood than control workers. Foreign workers'' reproductive decisions were not affected by the resident queen, their level of fertility being similar whether or not the queen was removed from the host colony. Workers'' physiological state at the time of introduction is also of crucial importance, since infertile workers failed to develop a reproductive phenotype in a foreign nest. Therefore, both internal and environmental factors appear to condition individual reproductive strategies in this species, suggesting that more complex decision-making mechanisms are involved in the regulation of worker reproduction than previously thought.     

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.     

A Route to Direct Fitness: Natural and Experimentally Induced Queen Succession in the Tropical Primitively Eusocial Wasp <Emphasis Type="Italic">Ropalidia marginata</Emphasis>

Insect societies are hallmarks of cooperation because one or a few queens monopolize reproduction and several non-reproductive workers cooperatively raise brood. However, the loss of the queen exposes a colony to potential reproductive conflict, which is resolved only after a new queen takes over. We studied queen succession in natural and experimental colonies of the primitively eusocial wasp Ropalidia marginata to understand the proximate behavioral strategies involved in the resolution of this conflict. Previous work has shown that in this species, experimental queen removal always results in only one worker becoming hyper-aggressive and taking over the colony as its next queen, without ever being challenged. Here we show that even during natural queen turnover, one and only one worker becomes hyper-aggressive and takes over as the next queen, without being challenged. During natural queen turn-over, aggression of the successor may sometimes begin before the loss of the old queen and may sometimes decline more rapidly, unlike in the case of experimental queen removal. The successor begins to lay eggs sooner after a natural queen turn-over as compared to experimental queen removal. This is expected because workers might detect the gradual decline of the queen preceding her disappearance. Because queen succession is expected to be more prevalent in tropical perennial species, we expect natural selection to have favored such an orderly queen succession so that a route to direct fitness is available without significant reduction in cooperation.     

Substantial direct fitness gains of workers in a highly eusocial ant

Hamilton's theory of inclusive fitness suggests that helpers in animal societies gain fitness indirectly by increasing the reproductive performance of a related beneficiary. Helpers in cooperatively breeding birds, mammals and primitively eusocial wasps may additionally obtain direct fitness through inheriting the nest or mating partner of the former reproductive. Here, we show that also workers of a highly eusocial ant may achieve considerable direct fitness by producing males in both queenless and queenright colonies. We investigated the reproductive success of workers of the ant Temnothorax crassispinus in nature and the laboratory by dissecting workers and determining the origin of males by microsatellite analysis. We show that workers are capable of activating their ovaries and successfully producing their sons independently of the presence of a queen. Genotypes revealed that at least one fifth of the males in natural queenright colonies were not offspring of the queen. Most worker‐produced males could be assigned to workers that were unrelated to the queen, suggesting egg‐laying by drifting workers.     

Sex-ratio regulation: the economics of fratricide in ants

In many insect societies, workers can manipulate the reproductive output of their colony by killing kin of lesser value to them. For instance, workers of the mound-building Formica exsecta eliminate male brood in colonies headed by a single-mated queen. By combining an inclusive fitness model and empirical data, we investigated the selective causes underlying these fratricides. Our model examines until which threshold stage in male brood development do the workers benefit from eliminating males to rear extra females instead. We then determined the minimal developmental stage reached by male larvae before elimination in F. exsecta field colonies. Surprisingly, many male larvae were kept until they were close to pupation, and only then eliminated. According to our model, part of the eliminated males were so large that workers would not benefit from replacing them with new females. Moreover, males were eliminated late in the season, so that new females could no longer be initiated, because matings take place synchronously during a short period. Together, these results indicate that workers did not replace male brood with new females, but rather reduced total brood size during late larval development. Male destruction was probably triggered by resource limitation, and the timing of brood elimination suggests that males may have been fed to females when these start to grow exponentially during the final larval stage. Hence, the evolution of fratricides in ants is best explained by a combination of ecological, demographic and genetic parameters.     

The queen colony phase of vespine wasps (Hymenoptera, Vespidae)

Characteristics of queen nests of the Vespinae are reviewed including the number of cells and each brood stage in the mature nest, cell-building and egg-laying rates, length of each brood stage and the occurrence of brood cannibalism. Success rates of queen nests of the Vespinae are reviewed with their causes of failure including the effect of queen usurpation, and with particular reference to variation of queen behaviour or queen quality. Evidence for the differences between potentially successful and unsuccessful queen nests is given. Simulation models of queen nests are developed for potential successful and unsuccessful colonies to determine the extent of larval cannibalism, larval life extension and for unsuccessful queen nests, oophagy. The variation of cell building rate, the importance of ambient temperature and the fast development of the first batch of eggs are considered in relation to polistine studies. The causes of the variation of queen quality are unknown but could be related to the amount of food received as larvae or as adults before leaving the natal colony.     

The impact of caging the queens during the flow season on some biological activities of honeybee colonies

This study was achieved in a private apiary located in a banana farm in Sa El Hagar, Basioun, Gharbia, Egypt from August 15, 2019 to May 25, 2020, including the banana (Musa sp., Musaceae) flow season (August and September) and extend to Egyptian clover (Trifolium alexandrinum L., Fabaceae) flow season (May). The study aimed to evaluate the effect of confining the queen during the banana flow season on the brood rearing, honey yield, and activation of worker's ovaries. Also, we determined the negative impact of caging the queen during the banana flow season on the activity of the colony in brood rearing, storing pollen, and honey yield after releasing the queen on 5 October, extending to the next flow season in May. The obtained results showed that the honeybee colonies with the caged queen produced significantly more honey yield and less brood production than the free queen ones during the banana flow season. Also, the caging of the queen did not affect the colony strength after releasing the queen despite the partial development of the ovaries of some workers, but they did not lay eggs. In addition, releasing the queens suppressed the ovaries of the laying workers. It can be concluded that caging the queen during the banana flow season helps the colonies to produce more honey yield without effect on the colony strength after releasing the queen despite the ovaries development of few workers without egg-laying.     

Regulation of reproduction in the neotropical ponerine ant,Pachycondyla villosa

Workers in queenless groups of the antPachycondyla villosa engage in antennal boxing and biting and by these interactions establish social dominance hierarchies, in which several high-ranking individuals may lay eggs. We observed egg cannibalism by dominant workers. The presence of larvae negatively effects the number of worker-produced eggs in queenless colony fragments in two ways. First, larvae feed on the eggs, and second, workers lay fewer eggs when larvae are present. In queenright colonies, workers lay eggs at a low rate, which are eaten by the queen or fed to the brood, but apparently they do not compete aggressively for egg-laying.     

Are workers of Atta leafcutter ants capable of reproduction?

Summary. Workers of most eusocial Hymenoptera can produce sons after queen loss, which (posthumously) benefits the queen and increases worker inclusive fitness. However, the evolutionary loss of worker ovaries has occurred in several lineages, while workers in other taxa may be infertile despite having ovaries. Workers of Atta leafcutter ants only lay trophic eggs in queenright colonies. Although Atta colonies are commonly kept at universities, museums, and zoos, no reports of worker sons in orphaned colonies exist, suggesting that Atta workers are infertile. To explicitly test this, we created eleven orphaned laboratory nests of Atta cephalotes, A. sexdens, and A. colombica, and maintained them for 3–6 months after queen loss. Eight colonies did not produce any brood, but three nests produced 1–4 worker-derived male larvae and pupae. Microsatellite genotyping indicated that these were worker sons. However, all males were tiny (3.5–9 mm long) compared to normal queen sons (16 mm long), and would almost certainly be unable to mate. We also found reproductive eggs, but most of these had no yolk and were thus inviable. We conclude that Atta workers are not completely infertile, but that worker fertility is low compared to the sister genus Acromyrmex, where workers routinely produce normally-size males after queen loss in the laboratory. We hypothesize that worker reproduction in orphaned Atta field colonies is almost never successful because the last workers die before their sons can be raised to adulthood, but that the importance of worker-laid trophic eggs for queen feeding has precluded the evolutionary loss of worker ovaries. Received 17 January 2005; revised 12 September 2005; accepted 3 October 2005.     

The value of oviposition timing,queen presence and kinship in a social insect

Reproductive cooperation confers benefits, but simultaneously creates conflicts among cooperators. Queens in multi-queen colonies of ants share a nest and its resources, but reproductive competition among queens often results in unequal reproduction. Two mutually non-exclusive factors may produce such inequality in reproduction: worker intervention or queen traits. Workers may intervene by favouring some queens over others, owing to either kinship or queen signals. Queens may differ in their intrinsic fecundity at the onset of oviposition or in their timing of the onset of oviposition, leading to their unequal representation in the brood. Here, we test the role of queen kin value (relatedness) to workers, timing of the onset of oviposition and signals of presence by queens in determining the maternity of offspring. We show that queens of the ant Formica fusca gained a significantly higher proportion of sexuals in the brood when ovipositing early, and that the presence of a caged queen resulted in a significant increase in both her share of sexual brood and her overall reproductive share. Moreover, the lower the kin value of the queen, the more the workers invested in their own reproduction by producing males. Our results show that both kinship and breeding phenology influence the outcome of reproductive conflicts, and the balance of direct and indirect fitness benefits in the multi-queen colonies of F. fusca.     

Effect of queen excluders on ovary activation in workers of the Eastern honeybee Apis cerana

Honeybee workers generally refrain from personal reproduction when a queen is present. Workers discern the presence and fecundity of the queen via volatile pheromones that permeate throughout the colony. Pheromones are emitted both by the queen herself and by the brood that she produces. If pheromone production is disrupted, some workers initiate egg laying. The Eastern honeybee Apis cerana is unusual in that workers have high levels of ovary activation even in the presence of a queen. To investigate the effect of disruption to pheromone dispersal, we fitted three A. cerana colonies with vertical queen excluders, thus splitting the colonies into a half containing a queen and a half without a queen. We regularly sampled adult workers from both sides of the excluder for 3 weeks. We also sampled workers from three control colonies that did not contain excluders. We found a significant increase in worker ovary activation 3 days after addition of excluders, suggesting that the reduced dispersal of pheromones allowed some workers to become reproductively active. Workers attempted to rear queen cells on the queenless halves of all three colonies. Queen-rearing ceased on day 9, at which time no queen-laid brood remained on the queenless halves of the colonies. Ovary activation rates continued to climb until day 9 and then gradually began to decline. With the exception of one egg, we did not observe worker-laid brood on the queenless side of the colonies, suggesting that workers continued policing eggs laid by workers. We conclude that if the distribution of brood pheromone is impeded, workers prepare to supersede their queen, accompanied by high levels of worker ovary activation. However, because workers continue to police each other, high ovary activation does not result in worker-produced drones.     

Queen pheromone regulates egg production in a termite

In social insects, resource allocation is a key factor that influences colony survival and growth. Optimal allocation to queens and brood is essential for maximum colony productivity, requiring colony members to have information on the total reproductive power in colonies. However, the mechanisms regulating egg production relative to the current labour force for brood care remain poorly known. Recently, a volatile chemical was identified as a termite queen pheromone that inhibits the differentiation of new neotenic reproductives (secondary reproductives developed from nymphs or workers) in Reticulitermes speratus. The same volatile chemical is also emitted by eggs. This queen pheromone would therefore be expected to act as an honest message of the reproductive power about queens. In this study, we examined how the queen pheromone influences the reproductive rate of queens in R. speratus. We compared the number of eggs produced by each queen between groups with and without exposure to artificial queen pheromone. Exposure to the pheromone resulted in a significant decrease in egg production in both single-queen and multiple-queen groups. This is the first report supporting the role of queen pheromones as a signal regulating colony-level egg production, using synthetically derived compounds in a termite.     

Queen pheromones in Temnothorax ants: control or honest signal?

Background  

The division of reproductive labor among group members in insect societies is regulated by "queen pheromones". However, it remains controversial whether these are manipulative, i.e., actively suppress worker reproduction, or honestly signal the fertility status of the queen to which workers react in their own interest by refraining from laying eggs. Manipulative queen control is thought to lead to an evolutionary arms race between queens and workers, resulting in complex queen bouquets that diverge strongly among different populations and species. In contrast, honest signals would evolve more slowly and might therefore differ less strongly within and among species.     

Morphology of reproductive and trophic eggs and their controlled release by workers in Trigona (Tetragonisca) angustula llliger (Apidae, Meliponinae)

Abstract. To study the reproductive potential of workers in the stingless bee Trigona (Tetragonisca) angustula Illiger (Apidae, Meliponinae), we examined the morphological quality of their eggs. Worker-eggs were all placed on the inside of the upper wall of brood cells. Normally, such eggs are consumed by the queen and are therefore referred to as trophic. The provisioning and oviposition processes in queenright colonies are characterized by the occurrence of circular aggregations of workers, 'rosettes', around the broodcell opening immediately after the release of a worker-egg. The ovaries of 35% of these rosette workers contained mature, chorionated eggs. In a single worker ovary, always only one mature egg was found. Some of the eggs, dissected from the ovarioles, showed a reticulate chorion pattern. Worker-eggs which lacked this pattern were significantly bigger than patterned eggs. Scanning electron micrographs revealed that the patterned worker-eggs are similar in appearance to queen-eggs.
After a worker-egg had been removed experimentally from a broodcell, the same cell could be oviposited by a worker again. Light microscopic analysis revealed that all these worker-eggs lacked the reticulate chorion pattern and were very similar in their morphology. In a colony without a laying queen workers laid eggs which had a reticulate pattern on the chorion. Since these eggs developed into males, we assume that the pattern on the chorion is characteristic for reproductive eggs. We also assume that the queen prevents the release of reproductive eggs by the workers. However, she does not inhibit the development of this type of egg.     

The influence of social structure on brood survival and development in a socially polymorphic ant: insights from a cross‐fostering experiment

Animal societies vary in the number of breeders per group, which affects many socially and ecologically relevant traits. In several social insect species, including our study species Formica selysi, the presence of either one or multiple reproducing females per colony is generally associated with differences in a suite of traits such as the body size of individuals. However, the proximate mechanisms and ontogenetic processes generating such differences between social structures are poorly known. Here, we cross‐fostered eggs originating from single‐queen (= monogynous) or multiple‐queen (= polygynous) colonies into experimental groups of workers from each social structure to investigate whether differences in offspring survival, development time and body size are shaped by the genotype and/or prefoster maternal effects present in the eggs, or by the social origin of the rearing workers. Eggs produced by polygynous queens were more likely to survive to adulthood than eggs from monogynous queens, regardless of the social origin of the rearing workers. However, brood from monogynous queens grew faster than brood from polygynous queens. The social origin of the rearing workers influenced the probability of brood survival, with workers from monogynous colonies rearing more brood to adulthood than workers from polygynous colonies. The social origin of eggs or rearing workers had no significant effect on the head size of the resulting workers in our standardized laboratory conditions. Overall, the social backgrounds of the parents and of the rearing workers appear to shape distinct survival and developmental traits of ant brood.