Insect societies discriminate against foreigners to avoid exploitation. In ants, helper workers only accept individuals with the familiar chemical cues of their colony. Similarly, unfamiliar eggs may get rejected at their first appearance in the nest. We investigated egg acceptance mechanisms by introducing different types of foreign eggs into worker groups of the ant Camponotus floridanus. Workers from established colonies familiar with queen-laid eggs always accepted eggs from highly fecund queens, but worker-laid eggs only after exposure for several weeks. Workers naive to eggs only rejected worker-laid eggs once they had prior exposure to eggs laid by highly fecund queens, suggesting that prior exposure to such eggs is necessary for discrimination. The general acceptance of eggs from highly fecund queens, irrespective of previous worker egg exposure, suggests an innate response to the queen pheromone these eggs carry. Workers learned to accept queen-laid eggs from different species, indicating high flexibility in learning egg-recognition cues. In incipient colonies with queen-laid eggs that carry a weak queen pheromone, worker-laid eggs were more likely to get accepted than queen-laid eggs from a different species, suggesting that the similarity of egg-recognition cues between the two types of C. floridanus eggs increases acceptance.

Social insects can sense colony size-even without visual information in a dark environment. How they achieve this is yet largely unknown. We empirically tested a hypothesis on the proximate mechanism using ant colonies. In Diacamma colonies, the monogynous queen is known to increase the effort devoted to queen pheromone transmission behaviour (patrolling) as the colony grows, as if she perceives colony size. The negative feedback hypothesis assumes that, through repeated physical contact with workers, the queen monitors the physiological state (fertility) of workers and increases her patrolling effort when she encounters more fertile workers. Supporting this hypothesis, we found that the queen increased her patrolling effort in response to a higher ratio of fertile workers under the experimental condition of constant colony size. Furthermore, chemical analyses and bioassays suggested that cuticular hydrocarbons have queen pheromone activity and can mediate the observed queen-worker communication of fertility state. Such a self-organizing mechanism of sensing colony size may also operate in other social insects living in small colonies.

The little fire ant (LFA), Wasmannia auropunctata, is a serious invasive pest first reported on Hawaii Island in 1999, and has since spread and established itself across the island. LFA is considered one of the worst 100 invasive species and has significant ecological, agricultural, and public health impacts in invaded areas, which include much of the tropical New World. Although localized eradication efforts have proven successful, they are intensive and difficult to implement. Furthermore, LFA\'s high invasive-ability resists these control efforts in areas where the species is established and can re-infest treated areas. This research set out to determine whether LFA queens have a suppressant effect on new queen production in nests, as a first step in identifying a potential queen pheromone for LFA. A queen pheromone could offer a means to shutdown LFA reproductive capability, potentially by suppressing the production of new queens or inducing the execution of queens or queen-destined larvae. When queenless experimental nests and polygyne experimental nests were compared, six out of eight queenless nests successfully reared both new alate queens (2.25 queens/nest) and drones (3.63 drones/nest) to adulthood, whereas only three of eight polygyne nests reared sexual larvae that failed to develop to adulthood or even the pupal stage. These results suggest that dealate mature LFA queens suppress the production of new alate queens in LFA nests, and is the first evidence that LFA may utilize a queen pheromone.

The queen mandibular pheromone (QMP) identified from the honeybee is responsible for maintaining reproductive division of labour in the colony, and affects multiple behaviours. Interestingly, QMP inhibits reproduction not only in honeybee workers, but also in distantly related insect species such as fruit flies and bumblebees. This study examines whether QMP also affects worker reproduction in the common wasp Vespula vulgaris. Wasp workers were exposed to one of the following treatments: QMP, wasp queen pheromone (the hydrocarbon heptacosane n-C27), or acetone (solvent-only control). After dissecting the workers, no evidence that QMP inhibits development in V. vulgaris could be found. However, this study could confirm the inhibitory effect of the hydrocarbon heptacosane on ovary activation. The reason why non-social species such as the fruit fly and social species such as bumblebees and ants respond to the QMP, while the social wasp V. vulgaris does not, is unclear. The investigation of whether olfaction is key to sensing QMP in other insect species, and the detailed study of odorant receptors in other social insects, may provide insights into the mechanisms of response to this pheromone.

The reproductive altruism in social insects is an evolutionary enigma that has been puzzling scientists starting from Darwin. Unraveling how reproductive skew emerges and maintains is crucial to understand the reproductive altruism involved in the consequent division of labor. The regulation of adult worker reproduction involves conspecific inhibitory signals, which are thought to be chemical signals by numerous studies. Despite the primary identification of few chemical ligands, the action modes of primer pheromones that regulate reproduction and their molecular causes and effects remain challenging. Here, these questions were elucidated by comprehensively reviewing recent advances. The coordination with other modalities of queen pheromones (QPs) and its context-dependent manner to suppress worker reproduction were discussed under the vast variation and plasticity of reproduction during colony development and across taxa. In addition to the effect of QPs, special attention was paid to recent studies revealing the regulatory effect of brood pheromones. Considering the correlation between pheromone and hormone, this study focused on the production and perception of pheromones under the endocrine control and highlighted the pivotal roles of nutrition-related pathways. The novel chemicals and gene pathways discovered by recent works provide new insights into the understanding of social regulation of reproductive division of labor in insects.

The objective of our project was to identify the sex attractant pheromone of queens of the slave-making ant Polyergus samurai, which was characterized as a blend of methyl 6-methylsalicylate and methyl 3-ethyl-4-methylpentanoate. Both compounds were identified in volatiles collected from squashed heads of queens, and in field trials, a blend of the two compounds was highly attractive to males. In contrast, males were not attracted to a blend of methyl 6-methylsalicylate with 3-ethyl-4-methylpentanol. Thus, the pheromone blend of this species is analogous to that of species in the Polyergus lucidus complex from eastern North America, rather than that of the Polyergus breviceps complex from western North America and the European species P. rufescens. These results are discussed in the context of speciation within the genus.

Eusociality is characterized by the reproductive division of labor between two castes: fertile queens and largely sterile workers. Queen pheromones are known to influence worker behavior and reproductive physiology and are therefore key components in regulating complex eusocial behavior [1]. Recent studies indicate that cuticular hydrocarbons (CHCs) act as queen pheromones in various eusocial hymenopteran species [2-8]. However, almost all species investigated to date are highly eusocial and do not include extant transitory stages from solitary to eusocial behavior [9]. Indeed, primitively eusocial species, which largely lack morphologically distinct castes, are thought to control worker reproduction through the physical aggression of the queen rather than via pheromones [10-12]. Halictid or sweat bees exhibit a high variability of eusociality including solitary and facultatively eusocial species [9, 13-16]. However, the mechanisms controlling worker reproduction in these transitory species are unknown. The results of a recent correlative study based on caste-specific chemical profiles in various halictid bees of different social levels have revealed an overproduction of macrocyclic lactones in queens compared with workers [17]. Using chemical analyses and behavioral experiments in which we simulated below-ground nests of the primitively eusocial sweat bee Lasioglossum malachurum, we identified a queen pheromone and found that macrocyclic lactones, not CHCs, influence worker behavior and decrease ovarian activation in this species. Our data suggest that the evolution of queen pheromones is more complex than previously inferred from highly eusocial species and shed new light on the complexity of the evolution of queen pheromones.

In eusocial insects, the high cost of altruistic cooperation between colony members has favoured the evolution of cheaters that exploit social services of other species. In the most extreme forms of insect social parasitism, which has evolved multiple times across most social lineages, obligately parasitic species invade the nests of social species and manipulate the workforce of their hosts to rear their own reproductive offspring. As alien species that have lost their own sociality, these social parasites still face social challenges to infiltrate and control their hosts, thus providing independent replicates for understanding the mechanisms essential to social dominance. This review compares socially parasitic insect lineages to find general trends and build a hypothetical framework for the means by which social parasites achieve reproductive dominance. It highlights how host social organization and social parasite life history traits may impact the way they achieve reproductive supremacy, including the potential role of chemical cues. The review discusses the coevolutionary dynamics between host and parasite during this process. Altogether, this review emphasizes the value of social parasites for understanding social evolution and the need for future research in this area.

Social wasps encompass species displaying diverse social organization regarding colony cycle, nest foundation, caste differences (from none to significant dimorphism) and number of reproductive queens. Current phylogenetic data suggests that sociality occured independently in the subfamily Stenogastrinae and in the Polistinae+Vespinae clade. In most species, including those with the simplest social organization, colony reproduction is monopolised by a single or few females. Since their nest mates can also develop ovaries and lay eggs, dominant females must somehow inhibit them from reproducing. Physical interactions in the form of open aggression or, usually, ritualised dominance by the fertile females contribute to fertility inhibition in several species, but it is unlikely to function in large colonies. In the latter case, reproduction within the colony is likely to be regulated through pheromones. Relatively little is known about these semiochemicals. Studies on all the three social wasp subfamilies, revealed that cuticular hydrocarbon components differ in abundance between egg-laying and not egg-laying females and that their composition depends on fertility status. In several species, females have been reported to manifestly react towards females with activated ovaries, but there is little evidence to support the hypothesis that fertile individuals are either recognized through their CHC composition, or that over-represented CHC constituents can inhibit fertility. Moreover, very little information exists on the possibility that exocrine glands release fertility signals or chemicals inhibiting fertility.

Social insects are well known for their extremely rich chemical communication, yet their sex pheromones remain poorly studied. In the thermophilic and thelytokous ant, Cataglyphis cursor, we analyzed the cuticular hydrocarbon profiles and Dufour\'s gland contents of queens of different age and reproductive status (sexually immature gynes, sexually mature gynes, mated and egg-laying queens) and of workers. Random forest classification analyses showed that the four groups of individuals were well separated for both chemical sources, except mature gynes that clustered with queens for cuticular hydrocarbons and with immature gynes for Dufour\'s gland secretions. Analyses carried out with two groups of females only allowed identification of candidate chemicals for queen signal and for sexual attractant. In particular, gynes produced more undecane in the Dufour\'s gland. This chemical is both the sex pheromone and the alarm pheromone of the ant Formica lugubris. It may therefore act as sex pheromone in C. cursor, and/or be involved in the restoration of monogyny that occurs rapidly following colony fission. Indeed, new colonies often start with several gynes and all but one are rapidly culled by workers, and this process likely involves chemical signals between gynes and workers. These findings open novel opportunities for experimental studies of inclusive mate choice and queen choice in C. cursor.