Brood adoption in the leaf-cutting ant <Emphasis Type="Italic">Acromyrmex echinatior</Emphasis>: adaptation or recognition noise?

The ability to discriminate between nestmates and non-nestmates is an important prerequisite for the evolution of eusociality. Indeed, social insect workers are typically able to discriminate between nestmate and non-nestmate workers. Adult non-nestmate workers are readily detected and rejected from the colony. Whether social insects can discriminate between nestmate and non-nestmate brood, however, is less clear. Here, we show that workers of the leaf-cutting ant Acromyrmex echinatior discriminate between nestmate and non-nestmate brood, and among brood of different stages. Initially, non-nestmate brood is attacked, but it is adopted after a delay. Adoption could occur due to inefficiency of the recognition system, or it could be adaptive because it is an inexpensive way to increase the workforce. Our results suggest that brood adoption may occur accidentally. We also report how workers replace fungal hyphae on the brood’s surface before transporting the brood into their fungus garden.     

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.     

A sensitive and reliable assay for queen discrimination ability in laboratory-reared workers of the ant Camponotus japonicus

The queen discrimination abilities of laboratory-reared Camponotus japonicus workers were examined individually by allowing them to carry their nestmate larvae toward either the mother queen or an alien queen. Source colonies had been reared under controlled conditions from founding queens and maintained at small size (< or = 10 workers each). Fifty-two of fifty-four workers raised in these eight different colonies carried nestmate larvae to the mother queen, and never carried them to the alien queen. Most of them attended nestmate larvae but never alien larvae. These results clearly demonstrate that the tested workers discriminate the nestmate queen and larvae from non-nestmate conspecifics. The assay used in this study is novel and sensitive, and may be suitable for neuroethological and molecular studies of social discrimination mechanisms.     

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.     

Differences in nestmate recognition for drones and workers in the honeybee, Apis mellifera (L.)

Nestmate recognition is the basic mechanism for rejecting foreign individuals and is essential for maintaining colony integrity in insect societies. However, in honeybees, Apis mellifera, both workers and males occasionally gain access to foreign colonies in spite of nest guards (=drifting). Instead of conducting direct behavioural observations, we inferred nestmate recognition for males and workers from the genotypes of naturally drifting individuals in honeybee colonies. We evaluated the degree of polyandry of the resident queens, because nestmate recognition theory predicts that the genotypic composition of insect colonies may affect the recognition precision of guards. Workers (N=1346) and drones (N=407) from 38 colonies were genotyped using four DNA microsatellite loci. Foreign bees were identified by maternity testing. The proportion of foreign individuals in a host colony was defined as immigration. Putative mother queens were identified if a queen's genotype corresponded with the genotype of a drifted individual. The proportion of a colony's individuals in the total number of drifted individuals was defined as emigration. Drones immigrated significantly more frequently than workers. The impact of polyandry was significantly different between drones and workers. Whereas drones immigrated more readily into less polyandrous colonies, worker immigration was not correlated with the degree of polyandry of the host colony. Furthermore, colonies with high levels of emigrated drones did not show high levels of emigration for workers, and colonies that adopted many workers did not adopt many foreign drones. Our data indicate that genetically derived odour cues are important for honeybee nestmate recognition in drones and show that different nestmate recognition mechanisms are used to identify drones and workers.     

Pavement Ant Workers (<Emphasis Type="Italic">Tetramorium caespitum</Emphasis>) Assess Cues Coded in Cuticular Hydrocarbons to Recognize Conspecific and Heterospecific Non-Nestmate Ants

Most ants live in closed societies from which non-members are excluded through fighting or ritualized displays to protect colony resources. Nestmate recognition is the process by which ants discriminate nestmate from non-nestmate ants. Ants use cues coded in mixtures of long-chain hydrocarbon compounds on the cuticle as nestmate recognition cues. Pavement ants (Tetramorium caespitum) form conspicuous wars between neighboring colonies that are organized after workers meet and make the decision to fight after assessing nestmate recognition cues. These wars involve thousands of individuals. Fighting is ritualized and few ants die in the process. We identified 24 cuticular hydrocarbon compounds, above 1% in relative abundance, in the profile of pavement ants with chain lengths ranging from 15 to 31 carbon atoms. Cuticular lipids contained, in order of abundance: mono-methyl alkanes (45–56%), n-alkanes (range: 16–40% relative abundance), and alkenes (10–20%), with small or trace amounts of di-methyl, tri-methyl alkanes and fatty acids. Results from behavioral tests show that pavement ants assess information in cuticular hydrocarbon profiles to recognize both conspecific and heterospecfic (Pogonomyrmex occidentalis and Camponotus modoc) non-nestmate ants and that the relative abundance of methyl-branched alkanes and alkenes codes for nestmate status, at least for conspecific interactions. Our data add to a growing body of knowledge about how ants use cuticular hydrocarbon based nestmate recognition cues to prevent the intrusion of non-nestmates in to colony space.     

A new ethological test to study nestmate recognition in adult ants

Summary An ethological test was designed to investigate nestmate recognition in adult ants. Contrary to classic dyadic tests, it consisted of a choice situation where a tested-worker was faced with nestmate and non-nestmate workers. These were kept alive, tied down, and thus immobilised in the neutral arena to record the tested-worker's reactions independently of the behaviour of the others. Such tests, applied to the ponerine ant,Ectatomma tuberculatum, enabled us to record various measures of discrimination and to limit aggressive behaviours between alien ants, which often leads to serious injury in dyadic tests. They also demonstrated the influence of age on discrimination behaviour in this species.     

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.     

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.

     

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.     

Role of Nest-paper Hydrocarbons in Nestmate Recognition of Dolichovespula maculata (L.) Workers (Hymenoptera: Vespidae)

The role of nest-paper hydrocarbons in nestmate recognition was examined in the social wasp Dolichovespula maculata. Pupae were excised from nine colonies of D. maculata and placed in separate gel capsules in the laboratory. When workers emerged, they were isolated in the absence of other wasps in one of four conditions: 1. With an untreated fragment of their natal nest; 2. With a fragment of their nest that had been extracted with hexane to remove surface hydrocarbons; 3. With a fragment of their nest that had been extracted with hexane, and had then had the extract containing the surface hydrocarbons reapplied; or 4. In the absence of any nest fragment. After 4 d, the newly emerged workers were tested for nestmate recognition with an experienced nestmate and an experienced non-nestmate in blind triplet tests. Prior exposure to nest-paper hydrocarbons had no significant effect upon the ability of newly emerged or experienced wasps to recognize their nestmates. Moreover, nestmate recognition did not occur in any treatment group.     

Nestmate discrimination in the harvester termite <Emphasis Type="Italic">Hodotermes mossambicus</Emphasis>

Summary Nestmate discrimination was studied in the African harvester termite Hodotermes mossambicus in trophallactic feeding experiments. The results show that Hodotermes mossambicus is able to discriminate nestmates from non-nestmates, feeding nestmates significantly more than non-nestmates. Experiments in which termites were treated with the antibiotic tetracycline for 48 hours and then used either as donors or as recipients in the trophallactic assay indicate that the degree of similarity in the intestinal flora strongly affects nestmate discrimination. Antibiotic-treated termites were fed more frequently by treated non-nestmates than by untreated nestmates. Pairs of non-nestmates, which were both treated with the antibiotic, exchange food as frequently as untreated nestmates. Pairs of nestmates, on the other hand, out of which only one animal was treated, exhibit nearly as little trophallactic contacts as the non-nestmate controls. The results parallel similar recent results in Reticulitermes speratus and indicate that termites can make use of recognition cues which are quite different from those of social hymenopterans, cues that are produced by intestinal symbionts.Received 5 November 2002; revised 17 and 27 February 2003; accepted 9 March 2003.     

The Effect of Queen Number on Nestmate Discrimination in the Facultatively Polygynous Ant Pseudomyrmex pallidus (Hymenoptera: Formicidae)

We present evidence indicating that the level of nestmate discrimination in the facultatively polygynous ant Pseudomyrmex pallidus varies predictably with the number of queens in the colony. P. pallidus workers were introduced into observation arenas in either nestmate or non-nestmate pairs. During the 5-min period immediately following the first contact between test ants, all interactions and relative distances were recorded. Aggression between non-nestmates was negatively correlated with the number of queens in the colony and distance between nestmates was positively correlated. These results are consistent with predictions of Reeve's (1989) optimal acceptance threshold model.     

Species and Colony Components in the Recognition Odor of Young Social Wasps: Their Expression and Learning (Polistes biglumis and P. atrimandibularis; Hymenoptera: Vespidae)

In Polistes, nestmate recognition relies on the learning of recognition cues from the nest. When wasps recognize nestmates, they match the template learned with the odor of the encountered wasp. The social wasp Polistes biglumis use the homogeneous odor of their colony to recognize nestmates. When these colonies become host colonies of the social parasite P. atrimandibularis, colony odor is no longer homogeneous, as the parasite offspring have an odor that differs from that of their hosts. In trying to understand how the mechanism of nestmate recognition works in parasitized colonies and why parasite offspring are accepted by hosts, we tested the responses of resident Polistes biglumis wasps from parasitized and unparasitized colonies to newly emerged parasites and to nestmate and non-nestmate conspecifics. The experiments indicate that immediately upon eclosion both young parasites and young hosts lack a colony odor and that colony odor can be soon acquired from the accepting colony. In addition, while residents of nonparasitized colonies recognize only the odor of their species, resident hosts of parasitized colonies have learned a template that fits the odors of two species.     

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.     

Social organization in some primitive Australian ants. I.Nothomyrmecia macrops Clark

Summary Results of laboratory-based ethological studies on twoNothomyrmecia macrops colonies with individually marked workers are reported. Interactive behavioural acts constituted less than 1% of all those recorded, revealing a strong tendency by the ants not to engage in social contact. Very few workers performed queen-directed acts. They stayed near the queen, though seldom in direct contact. Division of labour was otherwise barely apparent, except that some individuals showed a propensity to guard the nest entrance. No exchange of food was observed between workers, workers and queen, or adults and larvae (apart from worker placement of prey items with larvae). A queen fed from aDrosophila carcass retrieved from the nest floor, without assistance from workers. Systematic scanned observations confirmed levels of inactivity higher than previously observed in ants (comprising almost 2/3 of recorded behavioural acts). The time budget for activities directed toward the immature stages was the same in both colonies, and fluctuated during the circadian period. Non-nestmate larvae added to worker groups were more frequently licked than nestmate larvae, but this might not involve the particular recognition of nestmateversus non-nestmate brood. These observations support the hypothesis thatNothomyrmecia is primitively eusocial, and of special significance in myrmecology.     

The Effect of a Wasp's Age on Its Cuticular Hydrocarbon Profile and Its Tolerance by Nestmate and Non-Nestmate Conspecifics (Polistes fuscatus, Hymenoptera: Vespidae)

The effect of a wasp's age on its cuticular hydrocarbon profile and its tolerance by nestmate and non-nestmate conspecifics was investigated in a laboratory study of newly eclosed wasps ( Polistes fuscatus ) that were isolated from their comb at eclosion. In blind observations, mature females did not discriminate between young nestmates and non-nestmates that were ≤ 48 h-old. However, mature females did discriminate between young nestmates and non-nestmates that were 72 h old. Specifically, mature females were significantly more tolerant of (and significantly more likely to accept) 72 h-old nestmates than 72 h old non-nestmates. The abundance, relative abundance, and colony specificity of cuticular hydrocarbons changed significantly between 24 h-old and 72 h-old wasps. Our behavioral and chemical evidence indicates that wasps begin to develop a colony signature between 2 and 3 d of age.     

Genetic Analysis of the Drifting of Drones in Apis mellifera Using Multilocus DNA Fingerprinting

The presence of non-native drones in colonies of Apis mellifera was studied using multilocus DNA fingerprinting. Drones revealing a fingerprinting DNA banding pattern that did not correspond to the queen's genotype were classified as non-native animals. As previously reported for the drifting of workers, the position of the hive in relation to neighbouring colonies and the orientation of the flight entrance towards the sun showed significant correlations to the number of drifted drones in a colony. The frequency of non-native drones was low in central colonies (13 %) but high in marginal colonies (24 %). Furthermore, there was a significant correlation (r = 0.56) between the band-sharing coefficient of the non-native drones and the queen, and the number of drifted drones in the colony, which might indicate that genetically based nestmate recognition is involved in the drifting and/or acceptance of foreign drones in the colony.     

Spatial Aspects of Corpse Removal in the Western Harvester Ant, <Emphasis Type="Italic">Pogonomyrmex occidentalis</Emphasis>

Eusocial insects actively combat pathogen proliferation with a myriad of tactics, one of which is the removal of corpses from the nest, a behavior known as necrophoresis. Spatial patterns of corpse depositions by colonies of the western harvester ant, Pogonomyrmex occidentalis, were examined. Colonies were presented with nestmate and non-nestmate corpses to discern if each type of waste was handled differently. Specialized areas for corpse disposal were not observed. Non-nestmate corpses were carried farther from the nest than were nestmate corpses, perhaps reducing the chance of introduction of pathogens new to the colony that may be harbored by a non-nestmate. Factors external to the nest mound, such as slope and neighboring colonies, had no perceptible effect on these depositions and did not change the uniformity of dispersal of this particular waste. These findings add to our knowledge of the intricacies of corpse removal in eusocial insects and suggest that this is a more dynamic activity than previously thought.     

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.