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 body size of male honeybees (Apis mellifera L.) is a cause of skewed reproductive success. Large males are usually produced in colonies and have advantages in competition for mating and fertilisation. However, distinct small-sized males were produced depending on the colony conditions, particularly under queen-less conditions. Understanding the reproductive traits of small-sized males is currently limited, but it may provide insight into the developmental patterns and reproductive strategies that maximise reproductive success depending on body size and colony conditions. This study evaluated the process of sexual maturation in large- and small-sized males and the allometry between reproductive traits and body size. Changes in reproductive traits, including reproductive organs, number of spermatozoa, and sperm density, occurred earlier in small-sized males than in large-sized males after emergence. These results suggest that small males are precocious. The relatively early development of small-sized males would reflect the low developmental cost, which is likely to allow the production of many males and ensure reproductive success under circumstances in which available resources are limited. Furthermore, reproductive traits were positively correlated with body size, but allometry was different for these traits. Hence, the findings suggest that there is a given investment pattern toward reproductive traits with increasing body size, which would be responsible for high mating and fertilisation success in large males.

The modes through which individuals disperse prior to reproduction has important consequences for gene flow in populations. In honey bees (Apis sp.), drones (males) reproduce within a short flight range of their natal nest, leaving and returning each afternoon within a narrow mating window. Drones are assumed to return to their natal nests as they depend on workers to feed them. However, in apiaries, drones are reported to regularly make navigation errors and return to a non-natal nest, where they are accepted and fed by unrelated workers. If such a \"drone drift\" occurred in wild populations, it could facilitate some further degree of dispersal for males, particularly if drones drift into host nests some distance away from their natal nest. Here, we investigated whether drone drift occurs in an invasive population of the Asian honey bee (Apis cerana). Based on the genotypes of 1462 drones from 19 colonies, we found only a single drone that could be considered a candidate drifter (~0.07%). In three other colonies, drones whose genotypes differed from the inferred queen were best explained by recent queen turnover or worker-laying. We concluded that drone drift in this population is low at best, and A. cerana drones either rarely make navigation errors in wild populations or are not accepted into foreign nests when they do so. We therefore confirm that drone dispersal distance is limited to the distance of daily drone flights from natal nests, a key assumption of both colony density estimates based on sampling of drone congregation areas and population genetic models of gene flow in honey bees.

BACKGROUND: Worker reproduction has an important influence on the social cohesion and efficiency of social insect colonies, but its role in the success of invasive ants has been neglected. We used observations of 233 captive colonies, laboratory experiments, and genetic analyses to investigate the conditions for worker reproduction in the invasive Anoplolepis gracilipes (yellow crazy ant) and its potential cost on interspecific defence. We determined the prevalence of worker production of males and whether it is triggered by queen absence; whether physogastric workers with enlarged abdomens are more likely to be reproductive, how normal workers and physogastric workers compare in their contributions to foraging and defence; and whether worker-produced males and males that could have been queen- or worker-produced differ in their size and heterozygosity.
RESULTS: Sixty-six of our 233 captive colonies produced males, and in 25 of these, some males could only have been produced by workers. Colonies with more workers were more likely to produce males, especially for queenless colonies. The average number of days between the first appearance of eggs and adult males in our colonies was 54.1 ± 10.2 (mean ± SD, n = 20). In our laboratory experiment, queen removal triggered an increase in the proportion of physogastric workers. Physogastric workers were more likely to have yolky oocytes (37-54.9%) than normal workers (2-25.6%), which is an indicator of fertile or trophic egg production. Physogastric workers were less aggressive during interspecific aggression tests and foraged less than normal workers. The head width and wing length of worker-produced males were on average 4.0 and 4.3% greater respectively than those of males of undetermined source. Our microsatellite DNA analyses indicate that 5.5% of worker-produced males and 14.3% of males of undetermined source were heterozygous, which suggests the presence of diploid males and/or genetic mosaics in A. gracilipes.
CONCLUSIONS: Our experimental work provides crucial information on worker reproduction in A. gracilipes and its potential cost to colony defence. The ability of A. gracilipes workers to produce males in the absence of queens may also contribute to its success as an invasive species if intranidal mating can take place between virgin queens and worker-produced males.

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.

Kin selection and inclusive fitness are thought to be key factors explaining the reproductive altruism displayed by workers in eusocial insect species. However, when a colony\'s queen has mated with <2 males, workers may increase their fitness by producing their own male offspring. Conversely, when the queen has mated with ≥2 males, workers are expected to increase their inclusive fitness by eschewing the production of their sons and preventing other workers from reproducing as well. Here, we investigated sociogenetic structure and worker reproduction in the red honey ant, Melophorus bagoti. Morphometric analyses revealed that workers belong to one of two distinct subcastes: they are either majors or minors. Using DNA microsatellite markers, we showed that all the colonies had a single, multiple-mated queen and that there was no relationship between worker patriline and worker subcaste. Furthermore, we found that workers were producing males in the presence of the queen, which contrasts with the predictions of inclusive fitness theory. Although our results are based on a small sample, they can serve as the foundation for future research examining worker reproduction in M. bagoti.

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.

Workers of many species of social Hymenoptera have functional ovaries and are capable of laying haploid, unfertilized eggs, at least in the absence of a queen. Except for honeybees, it remains largely unknown whether worker-produced males have the same quality as queen-produced males and whether workers benefit in direct fitness by producing their sons. Previous studies in the monogynous ant Temnothorax crassispinus revealed that a high proportion of males in natural and laboratory colonies are worker offspring. Here, we compare longevity, body size, sperm length and sperm viability between queen- and worker-produced males. We either split queenright colonies into queenright and queenless halves or removed the queen from a fraction of the queenright colonies and then examined the newly produced males. Male quality traits varied considerably among colonies but differed only slightly between queen- and worker-produced males. Worker-produced males outnumbered queen-produced males and also had a longer lifespan, but under certain rearing conditions sperm from queen-produced males had a higher viability.

Resource inheritance is a major source of conflict in animal societies. However, the assumptions and predictions of models of conflict over resource inheritance have not been systematically tested within a single system. We developed an inclusive fitness model for annual eusocial Hymenoptera that predicts a zone of conflict in which future reproductive workers are selected to enforce nest inheritance before the queen is selected to cede the nest. We experimentally tested key elements of this model in the bumblebee Bombus terrestris. In colonies from which queens were sequentially removed, queen tenure was significantly negatively associated with worker male production, confirming that workers gain direct fitness by usurping the queen. In unmanipulated colonies, queen fecundity decreased significantly over the latter part of the colony cycle, confirming that workers\' indirect fitness from maintaining queens declines over time. Finally, in an experiment simulating loss of queen fecundity by removal of queens\' eggs, worker-to-queen aggression increased significantly and aggressive workers were significantly more likely to become egg layers, consistent with workers monitoring queen fecundity to assess the net benefit of future reproduction. Overall, by upholding key assumptions and predictions of the model, our results provide novel empirical support for kin-selected conflict over resource inheritance.

The hallmark of eusociality is the division of labour between reproductive (queen) and nonreproductive (worker) females. Yet in many eusocial insects, workers retain the ability to produce haploid male offspring from unfertilized eggs. The reproductive potential of workers has well-documented consequences for the structure and function of insect colonies, but its implications at the population level are less often considered. We show that worker reproduction in honey bees can have an important role in maintaining genetic diversity at the sex locus in invasive populations. The honey bee sex locus is homozygous-lethal, and, all else being equal, a higher allele number in the population lead to higher mean brood survival. In an invasive population of the honey bee Apis cerana in Australia, workers contribute significantly to male production: 38% of male-producing colonies are queenless, and these contribute one-third of all males at mating congregations. Using a model, we show that such male production by queenless workers will increase the number of sex alleles retained in nascent invasive populations following founder events, relative to a scenario in which only queens reproduce. We conclude that by rescuing sex locus diversity that would otherwise be lost, workers\' sons help honey bee populations to minimize the negative effects of inbreeding after founder events and so contribute to their success as invaders.