Honeybee (Apis cerana) guards do not discriminate between robbers and reproductive parasites

A hopelessly queenless honeybee colony has only one reproductive option: some workers must produce sons before the colony dies. This requires the workers to curtail egg policing (removal of worker-produced eggs), rendering the colony vulnerable to non-natal reproductive parasitism. In the Western honeybee, Apis mellifera, guarding (prevention of foreign workers from entering a colony) increases in queenless colonies, providing a defence against non-natal parasitism. However, in the closely related Eastern honeybee A. cerana, queenless colonies appear to be more tolerant of bees from other colonies. We presented guards of four A. cerana colonies with three types of workers: nestmate returning foragers, non-nestmate returning foragers and non-nestmates from a laying-worker colony. The latter are likely to have active ovaries, allowing us to test whether guard bees can detect which potential invaders are more likely to be reproductive parasites. After assessing guards’ reactions, we recaptured test bees and dissected them to determine levels of ovary activation. We found that nestmates were accepted significantly more frequently than the other two types of workers. However, there was no difference in the overall acceptance rates of non-nestmate returning foragers and bees from within laying-worker colonies. In addition, ovary-activated workers were no less likely to be accepted than those with inactive ovaries. Interestingly, colonies were more accepting of all three types of test bee after being made queenless. We conclude that, as has been previously suggested, guarding has no specific role in the prevention of non-natal parasitism in A. cerana.     

Specific recognition of reproductive parasite workers by nest-entrance guards in the bumble bee <Emphasis Type="Italic">Bombus terrestris</Emphasis>

Background

The impact of social parasites on their hosts’ fitness is a strong selective pressure that can lead to the evolution of adapted defence strategies. Guarding the nest to prevent the intrusion of parasites is a widespread response of host species. If absolute rejection of strangers provides the best protection against parasites, more fine-tuned strategies can prove more adaptive. Guarding is indeed costly and not all strangers constitute a real threat. That is particularly true for worker reproductive parasitism in social insects since only a fraction of non-nestmate visitors, the fertile ones, can readily engage in parasitic reproduction. Guards should thus be more restrictive towards fertile than sterile non-nestmate workers. We here tested this hypothesis by examining the reaction of nest-entrance guards towards nestmate and non-nestmate workers with varying fertility levels in the bumble bee Bombus terrestris. Because social recognition in social insects mainly relies on cuticular lipids (CLs), chemical analysis was also conducted to examine whether workers’ CLs could convey the relevant information upon which guards could base their decision. We thus aimed to determine whether an adapted defensive strategy to worker reproductive parasitism has evolved in B. terrestris colonies.

Results

Chemical analysis revealed that the cuticular chemical profiles of workers encode information about both their colony membership and their current fertility, therefore providing potential recognition cues for a suitable adjustment of the guards’ defensive decisions. We found that guards were similarly tolerant towards sterile non-nestmate workers than towards nestmate workers. However, as predicted, guards responded more aggressively towards fertile non-nestmates.

Conclusion

Our results show that B. terrestris guards discriminate non-nestmates that differ in their reproductive potential and respond more strongly to the individuals that are a greatest threat for the colony. Cuticular hydrocarbons are the probable cues underlying the specific recognition of reproductive parasites, with the specific profile of highly fertile bees eliciting the agonistic response when combined with non-colony membership information. Our study therefore provides a first piece of empirical evidence supporting the hypothesis that an adapted defensive strategy against worker reproductive parasitism exists in B. terrestris colonies.

     

Task Specialization and Odor Effects on Proboscis Extension Conditioning in Bumblebees (Bombus huntii)

Workers in a social insect colony have distinct experiences that may affect their performance in a learning task. In this study using free-foraging and flight-cage bumblebee Bombus huntii colonies, the strength of olfactory proboscis extension conditioning (PEC) was affected by a bee’s task specialization as a nurse or forager and the stimulus odor. Nurses (n?=?26) learned to respond to the odors 1-hexanol and lavender, but foragers (n?=?25) had inhibited conditioning to both odors. More nurses (73 %) than foragers (48 %) had at least one conditioned response (CR), and nurses displayed significantly more CRs than foragers. As expected, a pseudoconditioned control group (n?=?9) showed very few CRs. Among bees that were given a second day of trials, nurses—but not foragers—showed modest improvement. Such strong inhibition of PEC has not been described in honeybee or bumblebee foragers. The stimulus odor also affected conditioning strength in nurses. Lavender, a familiar odor, elicited earlier and more CRs than 1-hexanol. We propose that learning floral odors in the context of foraging may inhibit PEC in bumblebee foragers, whereas exposure to odors in the honey stores may prime subsequent learning in nurses.     

Social wasp parasites affect the nestmate recognition abilities of their hosts (<Emphasis Type="Italic">Polistes atrimandibularis</Emphasis> and <Emphasis Type="Italic">P. biglumis</Emphasis>, Hymenoptera,Vespidae)

Summary. Queens of the parasitic social wasp, Polistes atrimandibularis, temporarily mimic the odor of their host species, Polistes biglumis, but their offspring have parasite-specific odors. As a consequence, in parasitized colonies individuals with different odors co-inhabit the colony and host workers, who are responsible for colony defense, accept wasps with different odors. In order to verify whether this particular condition causes a change in recognition abilities of hosts, we tested nestmate/non-nestmate discrimination in field colonies invaded by social parasites (and in non-parasitized colonies as controls). Results show that parasitized colonies distinguish between nestmates and non-nestmates, distinguish their parasite queen from those that usurped alien colonies, and accept their parasite's non-mimetic offspring but make more recognition errors than non-parasitized colonies. The optimal acceptance threshold model predicts that when the frequency of encountering non-kin increases, residents become less permissive towards intruders. However, my data show that parasitized colonies are more permissive towards non-nestmates with respect to non-parasitized colonies but they are also more aggressive towards nestmates, suggesting that host workers' learning abilities are impaired.     

Intraspecific Food‐Robbing and Neighborhood Competition: Consequences for Anti‐Robber Vigilance and Colony Productivity

Most social animals have mechanisms to distinguish group members from outsiders, in part to prevent the exploitation of resources reserved for members of the group. Nevertheless, specialized thieves of the Neotropical ant, Ectatomma ruidum, also known as the ‘thieving ant’, regularly enter and steal resources from distinct, neighboring colonies. Here, we examine the mechanisms and consequences of thievery in a population of E. ruidum. We show that (1) individuals from nearby colonies were accepted more often than those from farther colonies; (2) rejection rates decreased as individuals interacted more with non‐nestmates from the same source colony; and (3) colonies that were experimentally treated to reduce thievery rates had improved productivity. The boost in productivity with thievery reduction was greater in low density populations than in high density populations. We conclude that, as in other species, thievery has negative fitness costs to E. ruidum. However, greater acceptance of neighbors than non‐neighbors and increased acceptance after habituation to non‐nestmates suggest a proximate explanation for the presence of thievery. Moreover, lower fitness costs of thievery at high nesting density, combined with observations of extraordinarily high densities of E. ruidum throughout its range, suggest there is little selection pressure among these ants to guard against thieves, thus providing an ultimate explanation why thievery persists among litter‐foraging ants.     

Chemical mimicry of the ant Oecophylla smaragdina by the myrmecophilous spider Cosmophasis bitaeniata: Is it colony-specific?

Workers of most social insects can distinguish between nestmates and non-nestmates, and actively attack the latter if they attempt to intrude into the nest or surrounding territory. Nevertheless, there are many records of heterospecific organisms living within the nests of social insects, and they are thought to gain access through chemical mimicry. The salticid spider Cosmophasis bitaeniata lives within the leaf nests of the ant Oecophylla smaragdina, where it preys on the ant larvae. We investigated, using behavioural bioassays and chemical analyses, whether the previously reported resemblance of the cuticular hydrocarbons of ant and spider was colony-specific. Behavioural experiments revealed that the spiders can distinguish between nestmate and non-nestmate major workers and are less inclined to escape when confined with ants that are nestmates. More significantly, C. bitaeniata were more likely to capture ant larvae from nestmate minor workers than non-nestmate minor workers. The chemical analyses revealed that the cuticular hydrocarbon profiles of the spiders and the major workers of the ant colonies were colony-specific. However, the hydrocarbon profiles of C. bitaeniata do not match those of the major workers of O. smaragdina from the same colony. Perhaps the colony-specific cuticular hydrocarbon profiles of C. bitaeniata function to obtain prey from the minor workers rather than avoid eliciting aggression from the major workers.     

Trophallaxis by temporal subcastes in the fire ant, Solenopsis invicta, in response to honey

ABSTRACT. Trophallactic behaviour of temporal subcastes in Solenopsis invicta Buren colonies was studied in two separate experiments. First, radiolabeled honey was presented to small colonies containing larvae, queens and marked workers; radioactivity in the head, thorax and abdomen of each ant was measured after 0.25, 1, 6, 12, 24, 48 and 72 h. The quantity of honey per subcaste was dependent on the total amount of honey collected. As more honey entered the colony, the quantity in foragers, queens and reserves increased more rapidly than that in nurses or larvae. The internal distribution of radioactivity in the head, thorax or abdomen indicated that differences between subcaste members in either the rate of food exchange or digestion existed. In a second experiment behavioural observations were made on marked workers in colonies before and after the presentation of honey. Reserves spent significantly more time engaged in trophallaxis than either foragers or nurses, both as active donors and active receivers. Based on number of encounters, nurses were more likely to receive honey while reserves and foragers were more likely to donate honey. The duration of encounters involving nurses tended to be shorter than those involving reserves or foragers.     

Kin recognition in the ant Rhytidoponera confusa II. Gestalt odour

Two models have been proposed for the transfer of genetically determined colony odour pheromones in social insects. The ‘Gestalt’ model suggests a complete transfer of pheromones amongst all nestmates whereas the ‘Individualistic’ model suggests no significant odour transfer with nestmates bearing individually distinct odours. In experiments with the ant Rhytidoponera confusa, colonies showed significantly more aggression (12% aggression) towards nestmates that had been kept in cages with non-nestmates than they did to nestmate controls (0% aggression). Colonies also showed significantly less aggression against non-nestmates that had been kept with nestmates in a separate cage (84% aggression) than they did to non-nestmate controls (100% aggression). A second experiment indicated that workers can absorb colony-specific odours from the nest materials of other colonies. However, whilst both experiments indicated a partial Gestalt component to the colony odour (ca. 28%), the Individualistic component seemed to be more important (ca. 72%).     

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.     

Food exchange behavior between multiple founding queens of Polyrhachis moesta (Hymenoptera: Formicidae) changes during hibernation

In some ant colonies founded by multiple queens, genetically unrelated queens cooperate to establish the colony. To test whether founding queens differentially adjust the amount of food exchange to nestmate and non-nestmate Polyrhachis moesta (Emery) queens before and after hibernation, food exchange in founding queens with different nutritional conditions was investigated. The proportion of non-nestmate queens showing food-begging behavior before hibernation was higher than that of nestmate queens. The number of begging behaviors was greater after hibernation than before hibernation, both between nestmates and between non-nestmates. Weight loss in fed queens performing food exchange with nestmate queens was significantly greater after hibernation than before hibernation. These results suggest that founding queens can discriminate between nestmate and non-nestmate founding queens and change the level of cooperation with founding queens before and after hibernation.     

Division of labour and specification of castes in the red imported fire ant Solenopsis invicta buren

Division of labour in Solenopsis invicta follows a familiar pattern: younger, smaller ants tend toward brood care while older, larger ants tend toward foraging. However, long-term observations of marked individuals reveal that length of nursing and foraging ‘careers’ and the age of transition between these activities vary considerably between and within size groups, and are related to length of life. Experiments with entire colonies show that larger ants are more likely than smaller ants to forage for insect prey. There are two main worker castes, ‘nurses’ and ‘foragers’, whose members span a wide age-size range, and a large ‘reserve’ subcaste, heterogeneous in age, size, and behaviour: reserves may nurse, forage, store liquid food, or relay food from nurses to foragers. The proportion of ants engaged in foraging decreases with colony size because many ants in large colonies are not exposed to recruitment signals.     

Nestmate recognition by guards of the Asian hive bee <Emphasis Type="Italic">Apis cerana</Emphasis>

When a honey bee colony becomes queenless and broodless its only reproductive option is for some of its workers to produce sons before the colony perishes. However, for this to be possible the policing of worker-laid eggs must be curtailed and this provides the opportunity for queenless colonies to be reproductively parasitized by workers from other nests. Such reproductive parasitism is known to occur in Apis florea and A. cerana. Microsatellite analyses of worker samples have demonstrated that the proportion of non-natal workers present in an A. cerana colony declines after a colony is made queenless. This observation suggests that queenless A. cerana colonies may be more vigilant in repelling potentially parasitic non-natal workers than queenright colonies. We compared rates of nestmate and non-nestmate acceptance in both queenright and queenless A. cerana colonies using standard assays and showed that there is no statistical difference between the proportion of non-nestmate workers that are rejected in queenless and queenright colonies. We also show that, contrary to earlier reports, A. cerana guards are able to discriminate nestmate workers from non-nestmates, and that they reject significantly more non-nestmate workers than nestmate workers. Received 25 February 2008; revised 21 May 2008; accepted 25 June 2008.     

Nestmate recognition of the stingless beeTrigona (Tetragonula) minangkabau (Apidae: Meliponinae)

Nestmate recognition was studied in the Southeast Asian stingless beeTrigona (Tetragonula) minangkabau, a species in which worker oviposition has not been observed in queenright or queenless colonies. When conspecific non-nestmate foragers from queenright and queenless colonies were introduced to the observed colony, they were all rejected by guards. Foragers of a different species (Trigona (Tetragonisca) angustula) were also completely rejected. However, conspecific non-nestmate callows were accepted as often as were nestmate callows, although guards recognized the difference. Accepted non-nestmate callows exchanged food with guards equally as much as nestmate callows did.     

Near-infrared spectroscopy identifies the colony and nest of origin of weaver ants, <Emphasis Type="Italic">Oecophylla smaragdina</Emphasis>

The ability of social insects to differentiate between colony members and others is essential for the survival of the colony. It enables individuals to direct altruistic behavior towards colony mates, while protecting the colony from intruders. Colonies have a distinct chemical signature that facilitates colony-mate recognition. However, in large polydomous colonies, this signal is likely to be modified by factors unique to each nest. We demonstrate, using near-infrared spectroscopy (NIRS), that individual weaver ants, Oecophylla smaragdina, can be differentiated with respect to their colony and nest of origin. 76.5% of individuals from four colonies could be correctly assigned to their colony of origin; and 79.6% of individuals could be assigned to the correct nest (of two) within their colony. Despite the differences between nests within colonies, in most cases individuals from one nest were more similar to individuals from the other nest within the colony than they were to individuals from any nest outside the colony. Therefore, a distinctive colony identity is maintained despite differences between nests within colonies. We discuss the advantages of using NIRS as a faster and less expensive alternative to the analysis of cuticular hydrocarbons following extraction and identification with gas chromatography/mass spectroscopy. Received 26 November 2007; revised 22 January 2008; accepted 25 January 2008.     

Behaviour of worker subcastes in the fire ant, Solenopsis invicta, in response to proteinaceous food

ABSTRACT. The effects of division of labour on response behaviour to food in the red imported fire ant, Solenopsis invicta Buren, were examined to determine if caste members differ in amount of food taken, in rate of food transfer, or in internal distribution of food; and to see if food availability, time, or temporal subcaste pairing affect feeding behaviour. To measure differences in behaviour we fed radioiodinated albumin mixed with egg yolk to colonies containing larvae, queens, and (a) foragers and nurses, or (b) foragers and reserves, or (c) nurses and reserves. Samples were taken over a 72-h period and radioactivity in the head, thorax and abdomen of each worker was determined. There were significant differences between nurses, foragers and reserves in quantity of food consumed, rate of transfer, and internal distribution of radioactivity. These differences were related to their respective roles of foraging, food storage and transfer, and brood tending. The quantity of food taken per subcaste was dependent on the total amount of food in the colony, with transfer rates differing between subcastes as the quantity of food in the colony increased. The rate at which protein was transferred between subcastes was slower in the reserves than that in either foragers or nurses. Therefore, reserves may serve as a temporary store of protein for the colony.     

Workers’ Extra-Nest Behavioral Changes During Colony Fission in Dinoponera quadriceps (Santschi)

Ant colonies can reproduce by two strategies: independent foundation, wherein the queen starts a new colony alone, and dependent foundation, in which workers assist the queen. In the queenless species Dinoponera quadriceps (Santschi), the colony reproduces obligatorily by fission, a type of dependent foundation, but this process is not well understood. This study describes a colony fission event of D. quadriceps in the field and analyzes the influence of the fission process on workers’ extra-nest behavior. Based on observations of workers outside the nest, five distinct stages were identified: monodomic stage, polydomic stage, split stage, conflict stage, and post-conflict stage. The colony was initially monodomic and then occupied a second nest before it split into two independent colonies, indicating a gradual and opportunistic dependent foundation. After the fission event, the daughter colony had aggressive conflicts with the parental colony, resulting in the latter’s disappearance. Colony fission affected workers’ extra-nest behavior by increasing the frequency of rubbing the gaster against the substrate (which probably has a chemical marking function) and by decreasing the frequency of foraging during the split stage. After the fission event, the number of foragers was halved and foragers remained nearer to the nest during extra-nest activity. The spatial closeness of the parental and daughter colonies led to competition that caused the extinction or migration of the parental colony. Intraspecific competition was indicated by foraging directionality at the colony level, whereby areas of neighbor colonies were avoided; this directionality was stronger while both colonies coexisted.     

Colony fusion and worker reproduction after queen loss in army ants

Theory predicts that altruism is only evolutionarily stable if it is preferentially directed towards relatives, so that any such behaviour towards seemingly unrelated individuals requires scrutiny. Queenless army ant colonies, which have anecdotally been reported to fuse with queenright foreign colonies, are such an enigmatic case. Here we combine experimental queen removal with population genetics and cuticular chemistry analyses to show that colonies of the African army ant Dorylus molestus frequently merge with neighbouring colonies after queen loss. Merging colonies often have no direct co-ancestry, but are on average probably distantly related because of overall population viscosity. The alternative of male production by orphaned workers appears to be so inefficient that residual inclusive fitness of orphaned workers might be maximized by indiscriminately merging with neighbouring colonies to increase their reproductive success. We show that worker chemical recognition profiles remain similar after queen loss, but rapidly change into a mixed colony Gestalt odour after fusion, consistent with indiscriminate acceptance of alien workers that are no longer aggressive. We hypothesize that colony fusion after queen loss might be more widespread, especially in spatially structured populations of social insects where worker reproduction is not profitable.     

The cuticular hydrocarbons profiles in the stingless bee Melipona marginata reflect task-related differences

Members of social insect colonies employ a large variety of chemical signals during their life. Of these, cuticular hydrocarbons are of primary importance for social insects since they allow for the recognition of conspecifics, nestmates and even members of different castes. The objectives of this study were (1) to characterize the variation of the chemical profiles among workers of the stingless bee Melipona marginata, and (2) to investigate the dependence of the chemical profiles on the age and on the behavior of the studied individuals. The results showed that cuticular hydrocarbon profiles of workers were composed of alkanes, alkenes and alkadienes that varied quantitatively and qualitatively according to function of workers in the colony.     

The guard honey bee: ontogeny and behavioural variability of workers performing a specialized task

Guarding is a relatively unstudied aspect of honey bee, Apis mellifera L., worker behaviour. The aim of this study was to characterize quantitatively the ontogeny and individual variability of guarding behaviour, the allocation of workers to the guard population in a colony, and the intercolonial variability of guarding behaviour. Guarding is a discrete task performed by a distinct group of workers that are younger than foragers and older than house bees. Workers that guarded initiated the behaviour between the ages of 7 and 22 days. The mean age of the onset of guarding varied; the minimum mean age of guards for a colony was 13·6 days and the maximum was 16·0 days. Workers varied in the length of time they spent as a guard. Most bees guarded for less than 1 days; however, some guarded up to 6 consecutive days. The more time a bee spent guarding during a day the more likely that bee was to guard for more than 1 day. Bees that guarded for more than 1 day also had longer and more frequent individual guarding bouts. All colonies that were studied had guard populations, but not all workers guarded. A relatively small proportion of any age cohort was observed to guard. The percentage of an age cohort that guarded varied among colonies, as did the size of the guard population. Guarding is a specialized task in that few bees guard, but guarding does not appear to require experience because so few bees remained as guards for very long. There was intercolonial variation in all aspects of the ontogeny of guarding and in allocation of workers to guarding. This variation is discussed in the light of other studies of variation in worker behaviour.     

Recognition in ants: social origin matters

The ability of group members to discriminate against foreigners is a keystone in the evolution of sociality. In social insects, colony social structure (number of queens) is generally thought to influence abilities of resident workers to discriminate between nestmates and non-nestmates. However, whether social origin of introduced individuals has an effect on their acceptance in conspecific colonies remains poorly explored. Using egg-acceptance bioassays, we tested the influence of social origin of queen-laid eggs on their acceptance by foreign workers in the ant Formica selysi. We showed that workers from both single- and multiple-queen colonies discriminated against foreign eggs from single-queen colonies, whereas they surprisingly accepted foreign eggs from multiple-queen colonies. Chemical analyses then demonstrated that social origins of eggs and workers could be discriminated on the basis of their chemical profiles, a signal generally involved in nestmate discrimination. These findings provide the first evidence in social insects that social origins of eggs interfere with nestmate discrimination and are encoded by chemical signatures.