Worker policing and worker reproduction in Apis cerana

Workers of the Asian hive bee, Apis cerana, are shown to have relatively high rates of worker ovary activation. In colonies with an active queen and brood nest, 1-5% of workers have eggs in their ovarioles. When A. cerana colonies are dequeened, workers rapidly activate their ovaries. After 4 days 15% have activated ovaries and after 6 days, 40%. A cerana police worker-laid eggs in the same way that A. florea and A. mellifera do, but are perhaps slightly more tolerant of worker-laid eggs than the other species. Nevertheless, no worker's sons were detected in a sample of 652 pupal males sampled from 4 queenright colonies. A cerana continue to police worker-laid eggs, even after worker oviposition has commenced in a queenless colony.     

Queen and worker policing in the tree wasp Dolichovespula sylvestris

Insect societies are sometimes exploited by workers who reproduce selfishly rather than help to rear the queens offspring. This causes a conflict-of-interest with the mother queen and, frequently, with the non-reproductive workers as well. One mechanism that can reduce conflict is policing, whereby either the queen or other workers aggress egg-laying workers or destroy worker-laid eggs. Here we present the first direct observations of queen and worker policing in natural, unmanipulated colonies of a social insect, the tree wasp Dolichovespula sylvestris. Worker reproduction was common, with workers producing 50% of all male eggs. However, most worker-laid eggs, 91%, were policed within 1 day, whereas most queen-laid eggs, 96%, remained unharmed. The workers were responsible for 51% of all policing events and the queen for 49%. The workers and mother queen also commonly aggressed ovipositing workers, and successfully prevented them from depositing eggs in 14% and 6% of all attempted ovipositions. Hence, both queen policing and worker policing occur and policing acts via two distinct mechanisms: selective destruction of worker-laid eggs and aggression of ovipositing workers. At a general level, our study shows that both centralized and decentralized control can act together to suppress conflict within social groups.Communicated by R. Page     

Co-occurrence of three types of egg policing in the Norwegian wasp <Emphasis Type="Italic">Dolichovespula norwegica</Emphasis>

In insect societies, workers often try to challenge the reproductive monopoly of the queen by laying their own eggs. Successful worker reproduction, however, is frequently prevented by queen policing or worker policing, whereby either the mother queen or non-reproductive workers selectively kill worker-laid eggs. Recently, a third mechanism—“selfish” worker policing—has also been described in which the workers selectively police worker-laid eggs but also lay eggs themselves. Here, we present results from the monogynous wasp Dolichovespula norwegica, which show that all three kinds of policing—queen policing, worker policing and “selfish” worker policing—co-occur. The net effect of these three kinds of policing collectively favoured the queen’s reproduction, as within 1 day 44% of the worker-laid eggs versus only 8% of the queen-laid eggs were eaten. Of the worker-laid eggs that were killed by workers, approximately two thirds were eaten by the reproductive workers even though these made up only a small proportion, 8%, of the work force. This means that policing workers obtained both direct fitness benefits as well as indirect (inclusive) fitness. In addition, we show that worker policing was carried out by a limited, specialised set of workers that was estimated to constitute approximately one quarter of the whole colony and of which 66% were non-reproductive.     

Changes in the cuticular hydrocarbons of incipient reproductives correlate with triggering of worker policing in the bulldog ant Myrmecia gulosa

In social insects, conflicts over male parentage can be resolved by worker policing. However, the evolution of policing behavior is constrained by the ability of individuals to identify reproductive nestmates, or their eggs. We investigated the occurrence of worker policing and its underlying chemical communication in the bulldog ant Myrmecia gulosa. Although workers have functional ovaries and can lay male-destined eggs, they do not reproduce in queenright colonies. To determine if their sterility is a consequence of worker policing, we experimentally induced worker reproduction in the presence of a queen. Some individuals were seized and immobilized by nestmates, and sometimes killed as a consequence. Although the ovarian development of immobilized individuals was variable, their cuticular hydrocarbon profiles were intermediate between reproductive and nonreproductive workers, indicating they were in the process of starting to reproduce. Approximately 29% of these incipient reproductive workers were successfully policed. To test for policing on eggs, we transferred viable worker eggs to queenright colonies and monitored their acceptance. Furthermore, we compared the surface hydrocarbons of the different types of eggs to determine whether these chemicals could be involved in egg recognition. We found that although there were differences in hydrocarbon profiles and discrimination between queen and worker-laid eggs, viable eggs were not destroyed. Our results strongly support the idea that cuticular hydrocarbons are involved in the policing of reproductive workers. A low level of worker policing appears sufficient to select for self-restraint in workers when few fitness benefits are gained by selfish reproduction. Policing of eggs may thus be unnecessary.     

The reproductive dilemmas of queenless red dwarf honeybee (Apis florea) workers

Honeybee (Apis) workers cannot mate, but retain functional ovaries. When colonies have lost their queen, many young workers begin to activate their ovaries and lay eggs. Some of these eggs are reared, but most are not and are presumably eaten by other workers (worker policing). Here we explore some of the factors affecting the reproductive success of queenless workers of the red dwarf honeybee Apis florea. Over a 2-year period we collected 40 wild colonies and removed their queens. Only two colonies remained at their translocated site long enough to rear males to pupation while all the others absconded. Absconding usually occurred after worker policing had ceased, as evidenced by the appearance of larvae. Dissections of workers from eight colonies showed that in A. florea, 6% of workers have activated ovaries after 4 days of queenlessness, and that 33% of workers have activated ovaries after 3 weeks. Worker-laid eggs may appear in nests within 4 days and larvae soon after, but this is highly variable. As with Apis mellifera, we found evidence of unequal reproductive success among queenless workers of A. florea. In the two colonies that reared males to pupation and which we studied with microsatellites, some subfamilies had much higher proportions of workers with activated ovaries than others. The significance of absconding and internest reproductive parasitism to the alternative reproductive strategies of queenless A. florea workers is discussed.     

Worker reproduction in honey-bees (Apis) and the anarchic syndrome: a review

Honey-bees, Apis, are an important model system for investigating the evolution and maintenance of worker sterility. The queen is the main reproductive in a colony. Workers cannot mate, but they can lay unfertilized eggs, which develop into males if reared. Worker reproduction, while common in queenless colonies, is rare in queenright colonies, despite the fact that workers are more related to their own sons than to those of the queen. Evidence that worker sterility is enforced by 'worker policing' is reviewed and worker policing is shown to be widespread in Apis. We then discuss a rare behavioural syndrome, 'anarchy', in which substantial worker production of males occurs in queenright colonies. The level of worker reproduction in these anarchic colonies is far greater than in a normal queenright honey-bee colony. Anarchy is a counterstrategy against worker policing and an example of a 'cheating' strategy invading a cooperative system.     

Evolution of worker sterility in honey-bees (Apis mellifera): how anarchistic workers evade policing by laying eggs that have low removal rates

In anarchistic honey-bee colonies, many workers’ sons are reared despite the presence of the queen. Worker-laid eggs are normally eaten by other workers in queenright colonies. Workers are thought to discriminate between queen-laid and worker-laid eggs by the presence or absence of a queen-produced egg-marking pheromone. This study compared the survival of three classes of eggs (worker-laid eggs from anarchistic colonies, worker-laid eggs from non-anarchistic queenless colonies, and queen-laid eggs) in both queenright normal colonies and queenright anarchistic colonies, in order to test the hypothesis that anarchistic workers evade policing by laying more acceptable eggs. As expected, few worker-laid eggs from non-anarchistic colonies survived more than 2 h. In contrast, worker-laid eggs from anarchistic colonies had much greater acceptability, which in some trials equalled the acceptability of queen-laid eggs. Anarchistic colonies were generally less discriminatory than normal queenright colonies towards worker-laid eggs, whether these originated from anarchistic colonies or normal queenless colonies. This indicates that the egg-removal aspect of the anarchistic syndrome involves both worker laying of eggs with greater acceptability and reduced discriminatory behaviour of policing workers. Received: 19 July 1999 / Received in revised form: 3 November 1999 / Accepted: 20 November 1999     

“Selfish worker policing” controls reproduction in a Temnothorax ant

Animal societies, including those of humans, are under constant threat by selfish individuals, who attempt to enforce their own interests at the cost of the group. In the societies of bees, wasps, and ants, such individual selfishness can be prevented by “policing,” whereby workers or queens impede the reproduction of other individuals by aggression, immobilization, or egg eating. In this study, we report on a particular kind of reproduction control in the ant Temnothorax unifasciatus, which can be considered as a selfish act itself. We experimentally induced workers to lay eggs by dividing several colonies into two halves, one with and one without a queen. In queenless colonies, workers established rank orders by aggression and several top-ranking workers started to reproduce. Upon reunification, egg-laying workers mostly stopped behaving aggressively. They were neither attacked by the queen nor by random workers, but instead received infrequent, nondestructive, targeted aggression from a few workers, most of which became fertile when the queen was later removed. The introduction of differentially stained worker-laid and queen-laid eggs in queenright fragments did not lead to a selective removal of worker-laid eggs. Hence, there appears to be no collective worker policing in T. unifasciatus. Instead, reproduction appears to be controlled mostly through a few attacks from high-ranking workers, which, in this way, might attempt to selfishly increase their chances of future reproduction.     

Egg-laying,egg-removal,and ovary development by workers in queenright honey bee colonies

Summary The study investigates whether worker policing via the selective removal of worker-laid male eggs occurs in normal honey bee colonies with a queen. Queenright honey bee colonies were set up with the queen below a queen excluder. Frames of worker brood and drone comb were placed above the queen excluder. Daily inspections of the drone frames revealed the presence of a few eggs, presumably laid by workers, at a rate of 1 egg per 16000 drone cells. 85% of these eggs were removed within 1 day and only 2% hatched. Dissections of workers revealed that about 1 worker in 10000 had a fully developed egg in her body. These data show that worker egg-laying and worker policing are both normal, though rare, in queenright honey bee colonies, and provide further confirmation of the worker policing hypothesis.     

Role of esters in egg removal behaviour in honeybee (Apis mellifera) colonies

In queen-right honeybee colonies workers detect and eat the vast majority of worker-laid eggs, a behaviour known as worker policing. However, if a colony becomes permanently queen-less then up to 25% of the worker population develops their ovaries and lay eggs, which are normally reared into a final batch of males. Ovary development in workers is accompanied by changes in the chemical secretion of the Dufour's gland with the production of queen-like esters. We show that ester production increases with the period that the colony is queen-less. The increased ester production also corresponds to an increase in persistence of worker-laid eggs in queen-right colonies, since the esters somehow mask the eggs true identity. However, in a rare queen-less colony phenotype, workers always eat eggs indiscriminately. We found that the egg-laying workers in these colonies were unusual in that they were unable to produce esters. This apparently maladaptive egg eating behaviour is also seen in queen-less colonies prior to the appearance of egg-laying workers, a period when esters are also absent. However, the indiscriminate egg eating behaviour stops with the appearance of ester-producing egg-laying workers. These observations suggest that esters are providing some contextual information, which affects the egg eating behaviour of the workers.     

Different policing rates of eggs laid by queenright and queenless anarchistic honey-bee workers (<Emphasis Type="Italic">Apis mellifera</Emphasis> L.)

Worker-reproduction is rare in queenright honey-bee colonies. When workers do lay eggs, their eggs are normally eaten by other workers presumably because they lack the queen's egg-marking signal. Workers use the absence of this queen signal to enforce the queen's reproductive monopoly by policing any worker-laid eggs. In contrast, in anarchistic colonies, the majority of the males arise from worker-laid eggs. Anarchistic worker-laid eggs escape policing because workers perceive anarchistic eggs as queen-laid. However, in this study, we show that eggs laid by queenless anarchistic workers do not escape policing and have very similar removal rates to worker-laid eggs from queenless wild-type (i.e. non-anarchistic) colonies. This suggests that, under queenless conditions, eggs laid by anarchistic workers lose their chemical protection and are therefore no longer perceived as queen-laid. Hence, the egg-marking signal seems to be only applied to eggs when queen and brood are present. This suggests that in the absence of queen and brood, the biosynthetic pathway that produces the egg-marking signal is switched off.Communicated by L. Keller     

Two independent mechanisms of egg recognition in worker <Emphasis Type="Italic">Formica fusca</Emphasis> ants

Informational constraints can be an important limitation on the accuracy of recognition. One potential constraint is the use of recognition information from the same sources in multiple discriminatory contexts. Worker wood ants, Formica fusca, discriminate eggs based on their maternal sources of origin in two main contexts: recognition of eggs laid by nestmate versus non-nestmate queens and recognition of worker-laid versus queen-laid eggs. We manipulated the experience of F. fusca workers in laboratory colonies to both worker-laid and queen-laid eggs by transferring eggs between colonies in order to investigate whether these two contexts of egg discrimination are independent. Experience of non-nestmate queen-laid eggs significantly increased worker acceptance of both familiar (18% accepted) and unfamiliar (10%) queen-laid eggs compared to control workers without experience of eggs other than those laid by their own colony’s queen (2%). In contrast, worker acceptance of worker-laid eggs was not affected by variation in the egg experience of workers (14% in workers from control colonies exposed only to eggs from their own colony’s queen versus 19% and 17% in workers from colonies which had received eggs laid by either a non-nestmate queen or nestmate workers, respectively). Our results suggest that these two recognition contexts do not strongly constrain each other and are different in their ontogeny. In particular, worker-laid eggs are universally discriminated against by workers from colonies with a queen whatever the egg experience of the workers, while non-nestmate queen-laid eggs are strongly discriminated against only by workers without experience of eggs laid by more than one queen.     

Queen and worker policing in the monogynous and monandrous ant, Diacamma sp.

Policing behavior that prevents workers from laying male eggs was examined in the monogynous and monandrous ponerine ant Diacamma sp. from Japan, in which a singly mated worker called a “gamergate” reproduces as the functional queen in each colony. Since oviposition by virgin workers is rare in the presence of a gamergate, we separated a portion of workers from the gamergates and induced their oviposition experimentally. When orphaned workers had started to oviposit, they were returned to the original colonies, where they continued to lay eggs for a while. The gamergates and other workers interfered with the laying workers by aggressively taking and finally eating the eggs. In total, 60% and 29% of the worker-derived eggs were eaten by gamergates and non-mother workers, respectively. The observed worker-worker interactions were not driven simply by competition to leave own sons, because non-laying non-orphaned workers interfered with worker reproduction. Furthermore, orphaned workers were usually attacked by non-orphaned workers soon after colony reunification. These results indicate that both queen policing by gamergates and worker policing in this species are mechanisms inhibiting worker oviposition. The gamergate contribution to policing was proportionately larger than that of workers, but among virgin workers, the relationship between dominance rank and contribution to policing was not clear. But about 11% of the eggs were not policed and were added to egg piles, especially in large colonies. Worker policing in a monandrous and monogynous eusocial Hymenoptera contrasts to other recent findings, and possible genetic, social, and ecological factors for its evolution in Diacamma sp. are discussed. Received: 16 November 1998 / Received in revised form: 9 February 1999 / Accepted: 21 February 1999     

Preservation and loss of the honey bee (<Emphasis Type="Italic">Apis</Emphasis>) egg-marking signal across evolutionary time

Honey bee workers are able to distinguish queen-laid eggs from worker-laid eggs, and remove (‘police’) worker-laid eggs. The cue that police workers use is as yet unidentified but is likely to be a chemical signal. This signal benefits queens for it ensures their reproductive monopoly. It also benefits collective workers because it allows them to raise more closely related queen-laid males than the less-related sons of half sisters. Because both parties benefit from the egg-marking signal, it should be stable over evolutionary time. We show that Apis mellifera workers can distinguish queen-laid from worker-laid eggs of the dwarf honey bee A. florea, a phylogenetically distant species that diverged from the A. mellifera lineage 6–10 mya. However, A. mellifera workers are unable to distinguish worker-laid eggs of A. cerana, a much more recent divergence (2–3 mya). The apparent change in the egg-marking signal used by A. cerana may be associated with the high rates of ovary activation in this species.     

Reproductive conflicts and egg discrimination in a socially polymorphic ant

The ability to discriminate against competitors shapes cooperation and conflicts in all forms of social life. In insect societies, workers may detect and destroy eggs laid by other workers or by foreign queens, which can contribute to regulate reproductive conflicts among workers and queens. Variation in colony kin structure affects the magnitude of these conflicts and the diversity of cues used for discrimination, but the impact of the number of queens per colony on the ability of workers to discriminate between eggs of diverse origin has so far not been investigated. Here, we examined whether workers from the socially polymorphic ant Formica selysi distinguished eggs laid by nestmate workers from eggs laid by nestmate queens, as well as eggs laid by foreign queens from eggs laid by nestmate queens. Workers from single- and multiple-queen colonies discriminated worker-laid from queen-laid eggs, and eliminated the former. This suggests that workers collectively police each other in order to limit the colony-level costs of worker reproduction and not because of relatedness differences towards queens’ and workers’ sons. Workers from single-queen colonies discriminated eggs laid by foreign queens of the same social structure from eggs laid by nestmate queens. In contrast, workers from multiple-queen colonies did not make this distinction, possibly because cues on workers or eggs are more diverse. Overall, these data indicate that the ability of F. selysi workers to discriminate eggs is sufficient to restrain worker reproduction but does not permit discrimination between matrilines in multiple-queen colonies.     

Worker rank, reproductive status and cuticular hydrocarbon signature in the ant, Pachycondyla cf. inversa

Workers in de-queened colonies of the neotropical ant, Pachycondyla cf. inversa, form linear or near-linear dominance hierarchies by violent antennation and biting. In these rank orders, social status and ovarian activity are on average highly correlated. Whereas the presence of a fertile queen appears to be sufficient to prevent workers from laying eggs, fertile workers do not completely control reproduction by their nestmates, suggesting that workers are able to differentiate between an egg-laying queen and an egg-laying worker. Here we show that the composition of cuticular hydrocarbons of egg-laying workers is quantitatively and qualitatively different from that of non-laying workers and resembles the hydrocarbon blend of the queen but does not completely match it. Furthermore, using discriminant analysis, it was possible to distinguish workers with four different classes of ovarian development based only on their cuticular hydrocarbon profiles. Fertility-associated changes in cuticular hydrocarbons may play an important role in the behavioural regulation of reproduction in this ant.     

Dufour's gland secretion of the queen honeybee (Apis mellifera): an egg discriminator pheromone or a queen signal?

The role of Dufour's gland secretion as an egg discriminator pheromone was reevaluated by simultaneously exposing workers to two combs, one containing queen- or worker-laid eggs and the second containing treated or untreated worker-laid eggs. Treatments included extracts of Dufour's gland secretion as well as the synthetic esters that were identified in the secretion. Policing was clearly detected both in queenright and queenless colonies by the swift removal of worker, but not of queen eggs. However, neither the glandular secretion nor its synthetic ester constituents were able to protect worker-born eggs from policing. Treated worker eggs were removed significantly faster than queen eggs, and at the same rate as non-treated worker eggs. These results are not consistent with the hypothesis that the secretion serves as an egg-marking pheromone. Chemical analyses of the queen abdominal tips revealed the presence of Dufour's esters, indicating that the glandular secretion oozes out and spreads over the cuticle around the genital chamber. However, contamination while ovipositing may also explain the minute amounts of these esters that were detected on the egg surface. Dufour's gland caste-specific composition suggests that in queens it may constitute a signal that plays a role in queen-worker interactions. Attraction bioassays revealed that the queen secretion, but not that of workers, is very attractive to workers. When applied either on a glass slide or on another worker, a retinue formed around the "surrogate queen". We conclude that Dufour's gland secretion constitutes part of a complex queen signal that is the basis for the social integrity of the honeybee colony.     

Intracolonial behavioral variation in worker oviposition,oophagy, and larval care in queenless honey bee colonies

Summary Two experiments were performed to determine whether worker reproduction in queenless honey bee colonies is influenced by colony genetic structure. In Experiment 1, allozyme analyses of workers and worker-derived drone larvae revealed that in half the colonies, there were genotypic differences in worker egg-laying behavior (presumed to involve actual oviposition), but biases in drone production were not always consistent with biases in egg-laying behavior. In Experiment 2, allozyme analyses again revealed intracolonial differences in egg-laying behavior and in behavior patterns thought to involve oophagy and larval care. Data support the hypothesis of a genetic influence on this intracolonial behavioral variation. Differences in the genotypic distributions of worker-derived drones relative to workers engaged in oviposition behavior in queenless colonies may be a consequence of genetic variability for egg production or for treatment of eggs and larvae (possibly coupled with kin recognition), or both. Offprint requests to: G.E. Robinson     

Worker reproductive parasitism and drift in the western honeybee Apis mellifera

When a honeybee (Apis spp.) colony loses its queen and is unable to rear a new one, some of the workers activate their ovaries and produce eggs. When a colony has a queen (i.e., it is queenright) almost all worker-laid eggs are eaten, but when hopelessly queenless, the workers become more tolerant of worker-laid eggs and rear some of them to adult drones. This increased tolerance renders a queenless colony vulnerable to worker reproductive parasitism, wherein unrelated workers enter the colony and lay eggs. Here, we show that the proportion of unrelated (non-natal) workers significantly decreases after an Apis mellifera colony becomes queenless. The remaining non-natal workers are as likely to have activated ovaries as natal workers, yet they produce more eggs than natal workers, resulting in significantly higher reproductive success for non-natal workers. In a second experiment, we provided queenless and queenright workers with a choice to remain in their own colony or to join a queenless or queenright colony nearby. The experiment was set up such that worker movement was unlikely to be due to simple orientation errors. Very few workers joined another colony, and there was no preference for workers to drift into or out of queenless or queenright colonies, in accordance with the proportion of non-natal workers declining significantly after becoming queenless in the first experiment.     

Reproductive conflict in honey bees: a stalemate of worker egg-laying and policing

 Using electrophoretic markers, eggs laid by workers were identified in honey bee (Apis mellifera) colonies with a queen. Based on extrapolation, these represented about 7% of the unfertilized (male) eggs laid in the colonies. A very small proportion of workers (of the order of 0.01%) lay these eggs. Worker-laid eggs are rapidly removed, so that very few sons of workers are reared. Thus the reproductive cooperation in bee colonies is maintained by ongoing antagonistic interactions among the members of the colony, with worker laying and egg removal policing by other workers being relatively common. Received: 24 November 1995/Accepted after revision: 25 May 1996