Nestmate Workers Research Articles

Identification of a colony- and dose-dependent worker aggregation pheromone in the subterranean termite Reticulitermes virginicus

Collective behaviors of social insects are often regulated by pheromones. In subterranean termites, some workers forage for and exploit decaying wood for new food resources while forming tunnels from their nest. Colonizing new food resources requires workers to build and disinfect tunnels and chambers inside the nest and ingest decaying wood; therefore subterranean termite colonies should have mechanisms to establish and maintain groups of workers to perform these functions. Recently, an aggregation pheromone was identified in workers of the termite Reticulitermes speratus, which induces quick attraction of nestmate workers and prolonged aggregation to the site of attraction. In this study, we extended this work to another species of Reticulitermes and identified a worker aggregation pheromone in the termite R. virginicus. GC–MS analysis and bioassays demonstrated that this pheromone consists of 3-octanone, 3-octanol, and palmitic acid and shows a colony-specific, dose-dependent attractant response but not an arrestant response. Furthermore, these pheromone components were most likely emitted from the surface of the body. This suggests that aggregation pheromone composition and function differ significantly among termite species, even within the same genus. This study advances our understanding of the regulatory mechanisms of termite aggregation behavior.

Fire ants feed their nestmates with their own venom

Venom secretion is widely used by ants for disease control and more generally as an external surface disinfectant. Here we report evidence that Solenopsis invicta feed their nestmates with their own venom. Venom alkaloids were found in crops and midguts of ants at concentration levels that have previously been reported as effective against various pathogens. These venom alkaloids were found in midguts of the larvae, indicating that trophallaxis must be involved in the transfer of venom, since larvae do not produce alkaloids and they depend on workers to be fed. After the mating flight, the female alates shed their wings, burrow into the soil, and start new colonies. The new queen provided alkaloids to her first batch of larvae in the new colony. Since the crops of female alates contain venom alkaloids donated from their nestmate workers, the transfer of worker alkaloids to new generation occurred. After minim adult workers emerged, they took the role in providing venom to the larvae in the colony. Minim adult workers eventually died out and the normal workers became the venom donors in the colony. Although other functions may be possible, considering the well-known antimicrobial property of venom alkaloids and their detected concentration levels, venom in the digestive system is most likely used as an internal antibiotic by fire ants.

A High Soldier Proportion Encouraged the Greater Antifungal Immunity in a Subterranean Termite.

Termites possess a mighty social immune system, serving as one of the key obstacles to controlling them biologically. However, the dynamic mechanism coordinating the social immunologic defense and caste distribution of the termites remains elusive. This study used the Coptotermes formosanus Shiraki and an entomopathogenic fungus as a host–pathogen system and experimentally manipulated a series of groups with different caste compositions of workers and soldiers. Then, the impact of demography on the behavior and innate immunity of termites was explored by analyzing the fungus susceptibility of the respective caste, efficiencies, and caste preferences of sanitary care, as well as the expression of the immune genes and phenoloxidase activity. Overall, to ensure the general health and survival of a group, the infected workers were found to sacrifice their survivorship for maintaining the soldier proportion of the group. If soldier proportion was limited within a threshold, both the survivorship of the workers and soldiers were not significantly affected by the infection. Correspondingly, the infected group with a higher proportion of soldiers stimulated the higher efficiency of a non-caste-biased sanitary care of the workers to the nestmate workers and soldiers. Moreover, the innate immunities of the infected workers were found to be more intensely upregulated in the group with higher soldier proportions. This suggested that the adjustable non-caste-biased sanitary care and innate immunity of the workers would contribute to the flexibility of the worker–soldier caste ratio in C. formosanus. This study, therefore, enhanced our understanding of the functional adaptation mechanism between pathogen-driven social immunity and the demography of the termites.

Tragedy of the commons in Melipona bees revisited.

Melipona stingless bees display a paradoxical overproduction of queens, which are later eliminated by nest-mate workers. Mechanistically, it was suggested that the monoterpenoid geraniol deposited into newly provisioned cells by adult bees would cause larvae to develop into queens in Melipona beecheii. This system could be evolutionarily stable if many of these new queens were to leave the nest and parasitize other genetically unrelated colonies nearby, as was shown to occur in a congeneric species. Here, we use microsatellite markers to test whether queen overproduction could be a strategy by which adult workers control the caste fate of the developing larvae to export copies of their own genes to the rest of the population via queen parasitism in M. beecheii. In addition, we re-examined whether artificially increasing the levels of geraniol indeed caused larvae to develop as queens rather than workers. Contrary to our prediction, we found no evidence for queen parasitism in M. beecheii and observed no effect of geraniol on the rearing of new queens. Together, these results support the original 'tragedy of the commons' hypothesis for queen overproduction in Melipona bees, where individual larvae selfishly bias their development towards the queen pathway according to their best evolutionary interests.

Oocyte resorption in termite queens: Seasonal dynamics and controlling factors

Female insects can resorb their oocytes that could not be oviposited. Oocyte resorption is proposed to be an adaptive mechanism to optimize fitness in hostile environments, recouping resources that might otherwise be lost. Social insects have developed reproductive division of labor, wherein a small number of queens are devoted to egg production. Matured queens are highly specialized in reproduction and are largely dependent on nestmate workers for their nourishment. Therefore, oocyte resorption in the queens should be influenced by social factors such as the amount of available workforce, as well as external and abiotic factors. In this study, we investigated the seasonal dynamics and regulation factors of oocyte resorption in actively reproducing termite queens. We continuously collected the field-nests of the subterranean termite Reticulitermes speratus and demonstrated that queens frequently resorbed their oocytes in late summer, even though it is one of the most productive seasons in this species. On the other hand, our laboratory experiment showed that oocyte resorption itself was strongly induced regardless of the season. We also found that the rate of oocyte resorption was influenced by colony size (the number of attending workers). These results suggest that termite queens seasonally resorb their oocytes, yet oocyte resorption itself is regulated by social factors rather than by seasonal factors. Our study provides a unique insight into the regulation of reproduction in social insects.

Transcriptomic analyses of the termite, Cryptotermes secundus, reveal a gene network underlying a long lifespan and high fecundity

Organisms are typically characterized by a trade-off between fecundity and longevity. Notable exceptions are social insects. In insect colonies, the reproducing caste (queens) outlive their non-reproducing nestmate workers by orders of magnitude and realize fecundities and lifespans unparalleled among insects. How this is achieved is not understood. Here, we identified a single module of co-expressed genes that characterized queens in the termite species Cryptotermes secundus. It encompassed genes from all essential pathways known to be involved in life-history regulation in solitary model organisms. By manipulating its endocrine component, we tested the recent hypothesis that re-wiring along the nutrient-sensing/endocrine/fecundity axis can account for the reversal of the fecundity/longevity trade-off in social insect queens. Our data from termites do not support this hypothesis. However, they revealed striking links to social communication that offer new avenues to understand the re-modelling of the fecundity/longevity trade-off in social insects.

Polyandry and paternity affect disease resistance in eusocial wasps

AbstractPolyandry (multiple mating by females) is a central challenge for understanding the evolution of eusociality. Several hypotheses have been proposed to explain its observed benefits in eusocial Hymenoptera, one of which, the parasite–pathogen hypothesis (PPH), posits that high genotypic variance among workers for disease resistance prevents catastrophic colony collapse. We tested the PPH in the polyandrous wasp Vespula shidai. We infected isolated workers with the entomopathogenic fungus Beauveria bassiana and quantified their survival in the laboratory. Additionally, we conducted a paternity analysis of the workers using nine microsatellite loci to investigate the relationship between survival and the matriline and patriline membership of the workers. As predicted by the PPH, nestmate workers of different patrilines showed differential resistance to B. bassiana. We also demonstrated variation in virulence among strains of B. bassiana. Our results are the first to directly support the PPH in eusocial wasps and suggest that similar evolutionary pressures drove the convergent origin and maintenance of polyandry in ants, bees, and wasps.

Emeryia, Xenometra, zoserka: it's a boy, again! The misleading morphology of Cardiocondyla male ants

Wingless “ergatoid” males of the ant genus Cardiocondyla have repeatedly been described as females of novel genera of workerless social parasites (Emeryia and Xenometra), which after recognition of their real nature were synonymized with Cardiocondyla. Examination of ants newly collected from Comoe National Park in Cote d’Ivoire, West Africa revealed that winged Cardiocondyla males have sparked a similar misidentification: the “winged female sexuals” of the supposed inquiline Cardiocondyla zoserka Bolton 1982, known only from the type material from Nigeria, are in fact the winged males of their presumed host, a species of the Cardiocondyla shuckardi group (sensu Seifert 2002). The types of C. zoserka are immediately recognizable by their strangely modified antennal funiculi with cup-shaped apical segments, which do not resemble any other known ant antenna. More than twenty winged individuals from two Cardiocondyla colonies from the flood plain of Comoe River exhibited the same strange modifications of their antennae and were otherwise also similar to the C. zoserka type specimens. However, these winged ants turned out to be winged males rather than winged female sexuals. The inspection of the holotype and one paratype of C. zoserka verified that they also were males: like the winged males from Comoe N.P., they have concealed male genitals and ocelli, which are considerably larger than those of the female sexuals of Cardiocondyla. All female sexuals and workers found in the colonies with the bizarre winged males from Comoe N.P. had antennae with a three-segmented club as is typical for this genus. Sequences of the mitochondrial cytochrome c oxidase subunits I (CO I) and II (CO II) of a winged male and two nestmate workers showed that they are close to C. venustula and C. shuckardi but differ from them in about 10% of the base pairs. Other samples of Cardiocondyla collected in Comoe N.P. belong to C. venustula and related species. Their males were always ergatoid or “intermorphic” (i.e., ergatoid with rudimentary wings), as previously found in other populations of C. venustula, and never winged with modified antennae.

Facing death: How does the subterranean termite Coptotermes gestroi (Isoptera: Rhinotermitidae) deal with corpses?

Social insects display a range of sophisticated behaviors to deal with cadavers, which together act to guarantee the health and social homeostasis of their colonies. In termites, cadaver management involves an entire repertoire of activities, such as antennation, grooming, retreat, alarm and agonistic behaviors. We performed different bioassays to evaluate whether postmortem-age, origin and caste of a cadaver influence the behavior of Coptotermes gestroi. Quantitative analyses of corpse management behaviors indicated that C. gestroi is able to discriminate the origin and caste of cadavers. Cadavers of worker nestmates were preferentially cannibalized while corpses of alien workers were buried. In addition, soldiers that had been dead for 24hours were buried while freshly dead soldiers were either buried, consumed or ignored, indicating a plastic behavioral response that depends on postmortem age. Corpse consumption is an important component of corpse management repertoire because this process eliminates contaminant and enables nutrient recycling. The physical isolation of the termite corpse through burial after grooming also helps to prevent other colony members from contacting the cadaver, further mitigating against the potential spread of disease.

The road to sociality: brood regulation of worker reproduction in the simple eusocial bee Bombus impatiens

Eusocial societies, in which egg laying is monopolized by one or a few females, have evolved multiple times during the evolution of insects but were always rooted in a simple family structure. Female reproduction in such families is often characterized by a trade-off between reproduction and brood care, yet most work on the regulation of reproduction in social insects has focused on signals and traits exhibited by adults: particularly on the behaviour and chemical signals produced by the queen and nestmate workers. Here we examined the role of brood in regulating worker reproduction in Bombus impatiens , an annual eusocial species whose reproduction is monopolized by the queen via an unknown mechanism. We found that the presence of young larvae reduced workers' egg laying, whereas the presence of pupae stimulated egg laying. The effect of young larvae was quantity dependent, with nearly complete suppression of egg laying in cages containing a pair of workers and more than 10 young larvae, and replicable regardless of worker age, relatedness to brood or brood parentage/sex. The findings that any larvae can regulate worker reproduction in this simple, yet eusocial, species highlight the role of brood in the evolution of advanced eusocial insects as a mechanism for regulating worker sterility. These findings also provide the first holistic explanation for the regulation of worker reproduction in B. impatiens , suggesting that the queen inhibits worker reproduction through her brood. • Presence of young larvae inhibited worker egg laying in B. impatiens . • Presence of pupae stimulated worker egg laying. • Larval effects were independent of relatedness, brood parentage or its sex. • Larval effect was quantity dependent. • Brood care plays an important role in the evolution of sociality.

Do Primitively Eusocial Wasps Use Queen Pheromones to Regulate Reproduction? A Case Study of the Paper Wasp Polistes satan

In several highly eusocial insect species with morphologically distinct castes, queen-characteristic cuticular hydrocarbons (CHCs) have been shown to act as queen signals that suppress the reproduction of nestmate workers. However, it is not known whether such queen pheromones might also play a role in regulating reproductive division of labour in primitively eusocial insects that lack morphologically defined castes. Here, we experimentally tested whether a blend of CHCs which occurred in higher concentrations in the dominant breeding female act as a queen pheromone, and inhibit reproduction by subordinate females in the primitively eusocial paper wasp Polistes satan. In contrast to earlier findings with highly eusocial species, our results show that although specific compounds were enhanced in dominant, reproductively active females, a blend of five of these compounds did not inhibit development of the ovaries of female nestmates. Instead, the dominant female had to be physically present to prevent subordinate females from reproducing. Our results are in line with earlier evidence suggesting that fertility-linked compounds in primitively eusocial wasps act only as cues and help to regulate reproduction when used in conjunction with aggressive dominance behaviour, meaning the physical presence of the dominant female. Alternatively, our results support the hypothesis that queen pheromones in highly eusocial insects were co-opted from fertility cues that were already present in primitively eusocial ancestors, even when initially, such compounds were merely produced as by-products of ovarian activation, without actually serving to signal reproductive status.

Supercolonial structure of invasive populations of the tawny crazy ant Nylanderia fulva in the US

BackgroundSocial insects are among the most serious invasive pests in the world, particularly successful at monopolizing environmental resources to outcompete native species and achieve ecological dominance. The invasive success of some social insects is enhanced by their unicolonial structure, under which the presence of numerous queens and the lack of aggression against non-nestmates allow high worker densities, colony growth, and survival while eliminating intra-specific competition. In this study, we investigated the population genetics, colony structure and levels of aggression in the tawny crazy ant, Nylanderia fulva, which was recently introduced into the United States from South America.ResultsWe found that this species experienced a genetic bottleneck during its invasion lowering its genetic diversity by 60%. Our results show that the introduction of N. fulva is associated with a shift in colony structure. This species exhibits a multicolonial organization in its native range, with colonies clearly separated from one another, whereas it displays a unicolonial system with no clear boundaries among nests in its invasive range. We uncovered an absence of genetic differentiation among populations across the entire invasive range, and a lack of aggressive behaviors towards conspecifics from different nests, even ones separated by several hundreds of kilometers.ConclusionsOverall, these results suggest that across its entire invasive range in the U.S.A., this species forms a single supercolony spreading more than 2000 km. In each invasive nest, we found several, up to hundreds, of reproductive queens, each being mated with a single male. The many reproductive queens per nests, together with the free movement of individuals between nests, leads to a relatedness coefficient among nestmate workers close to zero in introduced populations, calling into question the stability of this unicolonial system in which indirect fitness benefits to workers is apparently absent.

Cooperation and antagonism over time: a conflict faced by males of Tetragonisca angustula in nests

Before leaving to look for mates, the male stingless bees seem to face a dilemma to obtain food (energy) while avoiding aggression from the workers in their natal nests. It has been theorised that if these males waste time managing both situations inside the nests, it could reduce their chances of inseminating virgin queens outside nests. Here, I study Tetragonisca angustula (Hymenoptera: Apidae: Meliponini) males as a model of stingless bees to analyse whether the number of positive and negative interactions (cooperation = trophallaxis; antagonism = bites, respectively) with their worker nestmates are affected by the number of males inside nests or then by the time spent by them there. Next, I evaluate the binomial probability of these males leaving their natal nests as a function of trophallaxis and bites received of the workers. Finally, I assess which of the males’ body parts are commonly bitten by workers. The results demonstrate that males join with other males every day in hot places inside the nests to dehydrate nectar. Paradoxically, both trophallaxis and bites (these are often on legs) received from workers rise significantly throughout the time spent in the nests. Then, the probability of males leaving their natal nests rises 85% for every day spent inside the colonies. These data reinforce the suggestion of a dilemma faced by T. angustula males with relation to when to leave, i.e. how long to spend obtaining energy (and safety) within the nests while enduring aggression from workers, before permanently abandoning their natal nests to look for mates.

Effect of temperature and social environment on worker size in the ant Temnothorax nylanderi

Warm temperatures decrease insect developmental time and body size. Social life could buffer external environmental variations, especially in large social groups, either through behavioral regulation and compensation or through specific nest architecture. Mean worker size and distribution of worker sizes within colonies are important parameters affecting colony productivity as worker size is linked to division of labor in insect societies. In this paper, we investigate the effect of stressful warm temperatures and the role of social environment (colony size and size of nestmate workers) on the mean size and size variation of laboratory-born workers in the small European ant Temnothorax nylanderi. To do so, we reared field-collected colonies under medium or warm temperature treatments after having marked the field-born workers and removed the brood except for 30 first instar larvae. Warm temperature resulted in the production of fewer workers and a higher adult mortality, confirming that this regime was stressful for the ants. T. nylanderi ants followed the temperature size rule observed in insects, with a decreased developmental time and mean size under warm condition. Social environment appeared to play an important role as we observed that (i) larger colonies buffered the effect of temperature better than smaller ones (ii) colonies with larger workers produced larger workers whatever the rearing temperature and (iii) the coefficient of variation of worker size was similar in the field and under medium laboratory temperature. This suggests that worker size variation is not primarily due to seasonal environmental fluctuations in the field. Finally, we observed a higher coefficient of variation of worker size under warm temperature. We propose that this results from a disruption of social regulation, i.e. the control of nestmate workers over developing larvae and adult worker size, under stressful conditions.

Social regulation of ageing by young workers in the honey bee, Apis mellifera

Organisms' lifespans are modulated by both genetic and environmental factors. The lifespan of eusocial insects is determined by features of the division of labor, which itself is influenced by social regulatory mechanisms. In the honey bee, Apis mellifera, the presence of brood and of old workers carrying out foraging tasks are important social drivers of ageing, but the influence of young adult workers is unknown, as it has not been experimentally teased apart from that of brood. In this study, we test the role of young workers in the ageing of their nestmates. We measured the impact of different social contexts characterized by the absence of brood and/or young adults on the lifespan of worker nestmates in field colonies. To acquire insight into the physiological processes occurring under these contexts, we analyzed the expression of genes known to affect honey bee ageing. The data showed that young workers significantly reduced the lifespan of nestmate workers, similar to the effect of brood on its own. Differential expression of vitellogenin, major royal jelly protein-1, and methylase transferase, but not methyl farneosate epoxidase genes suggests that young workers and brood influence ageing of adult nestmate workers via different physiological pathways. We identify young workers as an essential part of the social regulation of ageing in honey bee colonies.

Location-specific cuticular hydrocarbon signals in a social insect.

Social insects use cuticular hydrocarbons (CHCs) to convey different social signals, including colony or nest identity. Despite extensive investigations, the exact source and identity of CHCs that act as nest-specific identification signals remain largely unknown. Perhaps this is because studies that identify CHC signals typically use organic solvents to extract a single sample from the entire animal, thereby analysing a cocktail of chemicals that may serve several signal functions. We took a novel approach by first identifying CHC profiles from different body parts of ants (Iridomyrmex purpureus), then used behavioural bioassays to reveal the location of specific social signals. The CHC profiles of both workers and alates varied between different body parts, and workers paid more attention to the antennae of non-nest-mate and the legs of nest-mate workers. Workers responded less aggressively to non-nest-mate workers if the CHCs on the antennae of their opponents were removed with a solvent. These data indicate that CHCs located on the antennae reveal nest-mate identity and, remarkably, that antennae both convey and receive social signals. Our approach and findings could be valuably applied to chemical signalling in other behavioural contexts, and provide insights that were otherwise obscured by including chemicals that either have no signal function or may be used in other contexts.

Defensive effect of soldier-specific secretion by Reticulitermes speratus (Isoptera: Rhinotermitidae) on the facultative termitophagous ponerine ant Brachyponera chinensis (Hymenoptera: Ponerinae)

Soldiers of the Japanese subterranean termite Reticulitermes speratus Kolbe possess the caste-specific terpenoid hydrocarbon β-selinene in the frontal gland. They secrete the compound as an alarm pheromone when facing danger. Furthermore, while dispersing the nestmate workers it simultaneously recruits the nestmate soldiers and enhances their aggression level. This study aims to confirm that the alarm pheromone might have defensive effects against facultative termite-hunting (termitophagous) ants, such as the Asian needle ant Brachyponera chinensis Emery. According to our behavioral bioassay, which provided fresh termite bodies, the ponerine ants used in the study immediately fed on the termite workers but avoided feeding on the soldiers. However, they avoided feeding on the workers when presented with the soldiers. Furthermore, extracted chemicals from the termite soldiers showed similar effects on the foraging activity of the ponerine ants, while those from the workers had no such effect. Soldier chemicals also clearly repelled the ponerine ants when they were treated at the entrance of a glass tube in which the worker bodies had been placed. It is most likely, therefore, that the termite soldier secretions serve as a chemical defense against the termitophagous ponerine ants.

No evidence of pre-copulatory mate choice by gynes in the facultatively parthenogenetic ant Cataglyphis cursor

Virgin queens (gynes) of the ant Cataglyphis cursor mate on the ground at the nest entrance, where they benefit from the protection of nestmate workers. By carrying out mating tests in the laboratory, we show that gynes mate with the first males they encounter, suggesting the absence of pre-copulatory mate choice by gynes. This is in agreement with the recent finding that workers behave aggressively towards foreign males and could thereby exercise a form of inclusive mate choice, which could replace pre-copulatory mate choice by gynes.

First Evidence of Inclusive Sexual Selection in the Ant Cataglyphis cursor: Worker Aggressions Differentially Affect Male Access to Virgin Queens

AbstractSexual selection typically shapes the traits of individuals capable of reproduction. However, in social species, it may also affect the behaviour of non‐reproductive individuals if this increases their inclusive fitness. We examined the potential role of workers during mating in the polyandrous ant Cataglyphis cursor. In this species, mating takes place on the ground near the nest entrance, and workers chase away foreign males trying to mate with young queens (gynes). We investigated the effects of worker aggression on male mating attempts by placing three or four males in the presence of one foreign gyne and 20 of her nestmate workers and contrasted this with a second group of 3–4 males in the presence of another gyne without nestmate workers (n = 8 paired replicates). Workers strongly attacked males (more than 20 aggressions per minute), although this did not affect the total number of mating attempts by the 3–4 males or the skew in the number of mating attempts among these males. However, there was an interaction between worker aggression and relative male weight, as relatively heavy males attempted to mate more often than lighter males in the absence of worker whereas the reverse occurred in the presence of workers. Our study is the first to experimentally test the potential role of non‐reproductive individuals in sexual selection. It shows that workers could potentially change male mating success by disfavouring heavier males.

Bumble bee workers drift to conspecific nests at field scales

1. Workers in several bee species travel to conspecific nests (‘drifting’), enter them, and produce male offspring inside them, so acting as intra‐specific social parasites. This adds a new dimension to bees' reproductive behaviour and spatial ecology, but the extent to which drifting occurs over field scales, i.e. at natural nest densities in field conditions, has been unclear.2. Using the bumble bee Bombus terrestris (Linnaeus) as a model system, we sought to determine rates of worker drifting at field scales and the frequency of potential drifter workers in wild nests.3. A field experiment with 27 colonies showed that workers travelled to, and became accepted in, conspecific nests that were up to 60 m away, although the number of accepted drifter workers within nests fell significantly with distance. The rate at which nests were entered by drifters was relatively high and significantly exceeded the rate at which drifters became accepted.4. Microsatellite genotyping of eight field‐collected nests from Greater London, U.K., showed that a low frequency (3%) of workers were not full sisters of nestmate workers and hence were likely to have been drifter workers.5. It is therefore concluded that workers can drift to conspecific nests over field scales and confirmed that successful drifting occurs in natural populations. Drifting appears to be a natural but low‐frequency behaviour permitting B. terrestris workers to gain direct fitness.