Chemical correlates of reproduction and worker policing in a myrmicine ant

In a number of wasps, bees, and in particular ponerine ants, quantitative and qualitative variation in the profile of cuticular hydrocarbons is associated with variation in fecundity and is likely to serve for communicating the reproductive status of an individual. Here, we demonstrate that the chemical profile on the cuticle of fertile workers and queens of the myrmicine ant Temnothorax unifasciatus is different from that of non-reproductive workers. Fertility and apparently also cuticular signatures are reversible under the influence of policing by worker aggression. Though no policing by egg-eating occurs in this species, queen and worker laid eggs also differed in their chemical profile.     

Individual recognition in ant queens

Personal relationships are the cornerstone of vertebrate societies, but insect societies are either too large for individual recognition, or their members were assumed to lack the necessary cognitive abilities . This paradigm has been challenged by the recent discovery that paper wasps recognize each other's unique facial color patterns . Individual recognition is advantageous when dominance hierarchies control the partitioning of work and reproduction . Here, we show that unrelated founding queens of the ant Pachycondyla villosa use chemical cues to recognize each other individually. Aggression was significantly lower in pairs of queens that had previously interacted than in pairs with similar social history but no experience with one another. Moreover, subordinates discriminated familiar and unfamiliar dominants in choice experiments in which physical contact, but not odor perception, was prevented and in tests with anaesthetized queens. The cuticular chemical profiles of queens were neither associated with dominance nor fertility and, therefore, do not represent status badges , and nestmate queens did not share a common odor. Personal recognition facilitates the maintenance of stable dominance hierarchies in these small societies. This suggests that the ability to discriminate between individual traits is selected for when it incurs net benefits for the resolution of conflict.     

Workers of a polistes paper wasp detect the presence of their queen by chemical cues

Differences in long-chain hydrocarbon mixtures among reproductive and nonreproductive individuals have been often revealed in social insects. However, very few papers demonstrated that these signatures actually act as contact pheromones used by nonreproductive to recognize the presence of a related queen in the colony. Cuticular and glandular hydrocarbons of Polistes paper wasps have been extensively studied, but, until now, the perception and recognition of such cues was not demonstrated. In this paper, we show, for the first time in Vespidae, that Polistes gallicus workers distinguish nestmates from alien individuals and queens from workers by the hydrocarbon mixtures of the Van der Vecht organ secretion (VVS). We also demonstrated that stroking behavior (a peculiar behavior of Polistes by which queens probably lay VVS on the nest) acts as an inhibitor of ovarian development in workers.     

Long-term memory of individual identity in ant queens

Remembering individual identities is part of our own everyday social life. Surprisingly, this ability has recently been shown in two social insects. While paper wasps recognize each other individually through their facial markings, the ant, Pachycondyla villosa, uses chemical cues. In both species, individual recognition is adaptive since it facilitates the maintenance of stable dominance hierarchies among individuals, and thus reduces the cost of conflict within these small societies. Here, we investigated individual recognition in Pachycondyla ants by quantifying the level of aggression between pairs of familiar or unfamiliar queens over time. We show that unrelated founding queens of P. villosa and Pachycondyla inversa store information on the individual identity of other queens and can retrieve it from memory after 24h of separation. Thus, we have documented for the first time that long-term memory of individual identity is present and functional in ants. This novel finding represents an advance in our understanding of the mechanism determining the evolution of cooperation among unrelated individuals.     

A role of the Dufour's gland in the dominance interactions of the paper wasp,Polistes fuscatus (Hymenoptera: Vespidae)

The Dufour's gland of the paper wasp, Polistes fuscatus,is a source of the cues used by dominant females to recognize the eggs laid by subordinates or nonnestmates on pre (worker)-emergence nests. When dominant wasps were presented with an egg covered with either (1) the Dufour's gland extract of a subordinate cofoundress, (2) the extract of an egg from the same subordinate, or (3) the solvent alone, the dominant female destroyed and replaced the eggs covered with the Dufour's extract significantly more frequently than the other eggs. Eggs with the extract of a nonnestmate's Dufour's gland were also eaten significantly more frequently than those with the solvent. Given similar choices, subordinates did not destroy any eggs. The Dufour's gland appears to have little or no role in communicating dominance directly among aggressively interacting cofoundresses.     

Visual signals of individual identity in the wasp Polistes fuscatus

Individual recognition is an essential component of interactions in many social systems, but insects are often thought incapable of the sophistication necessary to recognize individuals. If this were true, it would impose limits on the societies that insects could form. For example, queens and workers of the paper wasp Polistes fuscatus form a linear dominance hierarchy that determines how food, work and reproduction are divided within the colony. Such a stable hierarchy would be facilitated if individuals of different ranks have some degree of recognition. P. fuscatus wasps have, to our knowledge, previously undocumented variability in their yellow facial and abdominal markings that are intriguing candidates for signals of individual identity. Here, I describe these highly variable markings and experimentally test whether P. fuscatus queens and workers use these markings to identify individual nest-mates visually. I demonstrate that individuals whose yellow markings are experimentally altered with paint receive more aggression than control wasps who are painted in a way that does not alter their markings. Further, aggression declines towards wasps with experimentally altered markings as these novel markings become familiar to their nestmates. This evidence for individual recognition in P. fuscatus indicates that interactions between insects may be even more complex than previously anticipated.     

Cuticular hydrocarbons correlated with reproductive status in a queenless ant

Reproductive division of labour is regulated behaviourally in social insects lacking morphologically specialized castes. The directional nature of dominance interactions shows that recognition occurs, but little is known about its basis. In the queenless ant Dinoponera quadriceps, the top worker in the hierarchy (''alpha'') mates and produces offspring in each colony, while other workers remain virgin. Dominant ants frequently rub one antenna of subordinates against their own cuticle, and alpha and infertile nest-mates consistently differ in their relative proportions of the cuticular hydrocarbon 9-hentriacontene (9-C₃₁). The second-ranking ''beta'' occasionally lays unfertilized eggs and we show that she has less 9-C₃₁ than the alpha but more than infertile workers. To investigate further the link between 9-C₃₁ and ovarian activity, we experimentally removed alpha workers (n=11 individuals) and used solid-phase microextraction (SPME) with gas chromatography to measure changes in 9-C₃₁ on live beta workers which attained alpha status. The proportion of 9-C₃₁ on the replacement alpha increased significantly after six weeks, in parallel with her gain in fecundity. We discuss whether 9-C₃₁ provides honest information about egg-laying ability, enabling ants to recognize the different classes of nest-mates involved in reproductive conflicts. Such fertility cues could reliably underpin the antagonistic interactions occurring in insect societies.     

The look of royalty: visual and odour signals of reproductive status in a paper wasp

Reproductive conflicts within animal societies occur when all females can potentially reproduce. In social insects, these conflicts are regulated largely by behaviour and chemical signalling. There is evidence that presence of signals, which provide direct information about the quality of the reproductive females would increase the fitness of all parties. In this study, we present an association between visual and chemical signals in the paper wasp Polistes satan. Our results showed that in nest-founding phase colonies, variation of visual signals is linked to relative fertility, while chemical signals are related to dominance status. In addition, experiments revealed that higher hierarchical positions were occupied by subordinates with distinct proportions of cuticular hydrocarbons and distinct visual marks. Therefore, these wasps present cues that convey reliable information of their reproductive status.     

Nestmate recognition and identification of cuticular hydrocarbons composition in the swarm founding paper wasp Ropalidia opifex

Eusocial polistine wasps comprise species in which new colonies are founded by single wasps or groups of foundresses (independent founding species) or by swarms of queens and workers (swarm founding species). The first ones have relatively small societies generally comprising only one egg-laying queen, while the second ones have larger colonies where several egg-laying queens are usually present. These differences in social organisation are expected to influence the acceptance threshold of conspecifics in the colony and the intra-colony communication modalities. In this paper, we showed by field experiments that Ropalidia opifex (a swarm founding species) presents a good discrimination between alien and nestmate conspecifics. Moreover by GC/MS, we identified 19 cuticular hydrocarbons in this species, and we demonstrated that cuticular signature in this species depends mainly on colony membership, not showing any correlation with fertility.     

Endocrine mediated phenotypic plasticity: Condition-dependent effects of juvenile hormone on dominance and fertility of wasp queens

There has been increasing interest in the mechanisms that mediate behavioral and physiological plasticity across individuals with similar genotypes. Some of the most dramatic plasticity is found within and between social insect castes. For example, Polistes wasp queens can nest alone, dominate a group of cooperative queens, or act as worker-like subordinates who rarely reproduce. Previous work suggests that condition-dependent endocrine responses may play a role in plasticity between castes in the hymenoptera. Here, we test whether condition-dependent endocrine responses influence plasticity within castes in the wasp Polistes dominulus. We experimentally manipulate juvenile hormone (JH) titers in nest-founding queens and assess whether JH mediates variation in behavior and physiology. JH generally increased dominance and fertility of queens, but JH's effects were not uniform across individuals. JH had a stronger effect on the dominance and fertility of large individuals and individuals with facial patterns advertising high quality than on the dominance and fertility of small individuals and those advertising low quality. These results demonstrate that JH has condition-dependent effects. As such, they clarify how JH can mediate different behaviors in well nourished queens and poorly nourished workers. Many Polistes queens nest cooperatively with other queens, so condition-dependent hormonal responses provide a mechanism for queens to adaptively allocate energy based on their probability of successfully becoming the dominant queen. Research on the endocrine basis of plasticity often focuses on variation in endocrine titers alone. However, differential endocrine responses are likely to be a widespread mechanism mediating behavioral and physiological plasticity.     

Endocrine mediated phenotypic plasticity: Condition-dependent effects of juvenile hormone on dominance and fertility of wasp queens

There has been increasing interest in the mechanisms that mediate behavioral and physiological plasticity across individuals with similar genotypes. Some of the most dramatic plasticity is found within and between social insect castes. For example, Polistes wasp queens can nest alone, dominate a group of cooperative queens, or act as worker-like subordinates who rarely reproduce. Previous work suggests that condition-dependent endocrine responses may play a role in plasticity between castes in the hymenoptera. Here, we test whether condition-dependent endocrine responses influence plasticity within castes in the wasp Polistes dominulus. We experimentally manipulate juvenile hormone (JH) titers in nest-founding queens and assess whether JH mediates variation in behavior and physiology. JH generally increased dominance and fertility of queens, but JH's effects were not uniform across individuals. JH had a stronger effect on the dominance and fertility of large individuals and individuals with facial patterns advertising high quality than on the dominance and fertility of small individuals and those advertising low quality. These results demonstrate that JH has condition-dependent effects. As such, they clarify how JH can mediate different behaviors in well nourished queens and poorly nourished workers. Many Polistes queens nest cooperatively with other queens, so condition-dependent hormonal responses provide a mechanism for queens to adaptively allocate energy based on their probability of successfully becoming the dominant queen. Research on the endocrine basis of plasticity often focuses on variation in endocrine titers alone. However, differential endocrine responses are likely to be a widespread mechanism mediating behavioral and physiological plasticity.     

Rank integration in dominance hierarchies of host colonies by the paper wasp social parasite Polistes sulcifer (Hymenoptera, Vespidae)

In multiple-foundress nests of the wasp Polistes dominulus, dominance hierarchies are established among foundresses, and only the dominant (=alpha) individual lays eggs. The alpha female can be distinguished from subordinate females and workers on the basis of the proportions of some hydrocarbons present on the cuticle, suggesting that chemical signaling of her reproductive status could occur. P. dominulus is also the host species of the obligate social parasite Polistes sulcifer. After aggressively usurping host colonies and behaviorally replacing the host alpha female, parasites are characterized by a change in the proportions of their cuticular hydrocarbons to match that of the host cuticular profile at both species and colony levels. In the current study, we demonstrate that P. sulcifer queens also modify their cuticular hydrocarbon proportions after usurpation to match that of the host alpha female. Parasite females, therefore, acquire the dominant rank in host colonies both reproductively and chemically by mimicking the typical alpha profile of the host. Parasite females were not able to fully inhibit ovary development in host foundresses, and 10 days after usurpation, parasites, alpha and beta foundresses show similar chemical profiles and ovarian development.     

Orphaning does not affect the colony productivity of the primitive eusocial wasp Polistes snelleni

In colonies of primitively eusocial wasps, some dominant workers become successive queens and inherit queenship after the death of the foundress queens. Although workers in many species do not mate, workers of Polistes snelleni are capable of mating and female production. In this study, we removed foundress queens from colonies of P. snelleni to evaluate the effects of queen loss on the dominant–subordinate relationships among the remaining workers and the productivity of colonies in the species. The foundress queens were the sole egg layers in almost all of the queenright colonies. The frequency of dominance behaviour among the wasps in the queenright colonies was significantly less than in the orphan colonies. The frequency of dominance behaviour in the successive queens after queen removal was significantly more than in the foundress queens. Multiple workers had developed ovaries, including the successive queens in 66.7 % (10/15) of the orphan colonies after queen removal. The orphan colonies produced significantly more cells and eggs than the queenright colonies. Our results suggest that the reproductive potential of the successive queens in the orphan colonies is not lower than that of the foundress queens, and that the productivity of the orphan colonies is maintained rather than causing potential conflict over direct reproduction among workers.     

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.     

How to fool a virgin: Artificial dealation triggers oviposition in virgin Lasius niger queens

Once inseminated, ant queens rapidly shed their wings and start to lay eggs. Here we test whether there is a causal link between dealation and oviposition in the ant Lasius niger. We show that artificially dealated virgin queens start to lay eggs shortly after wing removal, whereas winged virgin queens hardly ever lay eggs. Dealate virgins do, however, produce fewer eggs than mated queens. These findings indicate that dealation does induce egg-laying, and that other factors, such as mating and/or insemination, further stimulate oviposition under natural conditions. Received 2 January 2006; revised 8 February 2006; accepted 4 April 2006.     

Cuticular hydrocarbons rather than peptides are responsible for nestmate recognition in Polistes dominulus

A colony of social insects is like a fortress where access is allowed only to colony members. The epicuticular mixture of hydrocarbons has been widely reported to be involved in nestmate recognition in insects. However, recent studies have shown that polar compounds (mainly peptides) are also present, mixed with hydrocarbons, on the cuticle of various insects, including the paper wasps of the genus Polistes. As these polar compounds are variable among Polistes species and are perceived by the wasps, this cuticular fraction could also be involved in nestmate recognition. Through MALDI-TOF (Matrix-Assisted Laser Desorption Ionization Time of Flight) mass spectrometry analysis, we assessed, for the first time, the intercolonial variability of the cuticular polar fraction of Polistes dominulus in order to evaluate its reliability as source of nestmate recognition cues. We then tested through behavioral assays the importance of the 2 isolated fractions (apolar and polar) in nestmate recognition by presenting them separately to colonies of P. dominulus. Our results showed that the cuticular polar compounds are not colony specific and they are not used by paper wasps to discriminate nestmates from non-colony members. On the contrary, we confirmed that the isolated cuticular hydrocarbons are the chemical mediators prompting nestmate recognition in paper wasps.     

Direct assessment of queen quality and lack of worker suppression in a paper wasp

Assessing a conspecific's potential is often crucial to increase one's fitness, e.g. in female choice, contests with rivals or reproductive conflicts in animal societies. In the latter, helpers benefit from accurately assessing the fertility of the breeder as an indication of inclusive fitness. There is evidence that this can be achieved using chemical correlates of reproductive activity. Here, we show that queen quality can be assessed by directly monitoring her reproductive output. In the paper wasp Polistes dominulus, we mimicked a decrease in queen fertility by regularly removing brood. This triggered ovarian development and egg-laying by many workers, which strongly suggests that brood abundance is a reliable cue of queen quality. Brood abundance can be monitored when workers perform regular brood care in small size societies where each brood element is kept in a separate cell. Our results also show that although the queen was not manipulated, and thus remained healthy and fully fertile, she did not control worker egg-laying. Nevertheless, when workers laid eggs, the queen secured a near reproductive monopoly by selectively destroying these eggs, a mechanism known as 'queen policing'. By contrast, workers destroyed comparatively few queen-laid eggs, but did destroy each other's eggs.     

The occurrence of sexual reproduction among ant workers

In less than 100 species of ponerine ants, queens no longer exist and have been replaced by mated egg-laying workers. Workers in other subfamilies can lay haploid eggs when queens are removed, but they never reproduce sexually. Ponerine workers are able to mate because they have a spermatheca in most species, foreign males are sexually active near their nests, and their pygidial gland secretions can assume a sexual meaning. Furthermore, ponerine queens are seldom very fecund, and one or several gamergates are able to approximate their egg production. Finally, opportunities for colony fragmentation occur consequent to their life history, and this is a necessary precondition because gamergates cannot start new colonies independently. Many of these characteristics are associated with the limited caste divergence exhibited in this phylogenetically primitive group. Although a few non-ponerine species exhibit some of these preconditions, gamergates have not been found outside the Ponerinae, which alone exhibit the combination of traits leading to queen elimination and worker mating.     

Rank and epicuticular hydrocarbons in different populations of the paper wasp <Emphasis Type="Italic">Polistes dominulus</Emphasis> (Christ) (Hymenoptera,Vespidae)

Summary Several studies of social insects have shown that epicuticular hydrocarbons are involved in recognition. The hypothesis is that the animals can use differences in chemical composition to acquire information about conspecific status (sex, colony, reproductive status or caste recognition).In this study, we searched for differences between the epicuticular profiles of alpha and beta co-foundresses in Polistes dominulus (Christ) colonies from three separate localities. Our aim was to identify specific compounds or sets of compounds which could be general indexes of foundress rank position in an associative foundation.We found quantitative and qualitative differences in the epicuticular profile among the three populations. The compounds that differentiate between alpha foundresses and their subordinates in the two neighbouring localities were not the same as in the third population. However, in all localities the alpha foundresses of each associative foundation presented a higher proportion of heavy compounds than their respective subordinates. Chemical differences related to rank position may be a fertility signal and are probably caused by physiological and behavioural characteristics, although it is not yet certain if they are actually used by the wasps.Received 28 November 2003; revised 22 February 2004; accepted 25 February 2004.     

Social context predicts recognition systems in ant queens

Recognition of group‐members is a key feature of sociality. Ants use chemical communication to discriminate nestmates from intruders, enhancing kin cooperation and preventing parasitism. The recognition code is embedded in their cuticular chemical profile, which typically varies between colonies. We predicted that ants might be capable of accurate recognition in unusual situations when few individuals interact repeatedly, as new colonies started by two to three queens. Individual recognition would be favoured by selection when queens establish dominance hierarchies, because repeated fights for dominance are costly; but it would not evolve in absence of hierarchies. We previously showed that Pachycondyla co‐founding queens, which form dominance hierarchies, have accurate individual recognition based on chemical cues. Here, we used the ant Lasius niger to test the null hypothesis that individual recognition does not occur when co‐founding queens do not establish dominance hierarchies. Indeed, L. niger queens show a similar level of aggression towards both co‐foundresses and intruders, indicating that they are unable of individual recognition, contrary to Pachycondyla. Additionally, the variation in chemical profiles of Lasius and Pachycondyla queens is comparable, thus informational constraints are unlikely to apply. We conclude that selection pressure from the social context is of crucial significance for the sophistication of recognition systems.