Kin discriminators in the eusocial sweat bee <Emphasis Type="Italic">Lasioglossum malachurum</Emphasis>: the reliability of cuticular and Dufour’s gland odours

The ability to discriminate degrees of relatedness may be expected to evolve if it allows unreciprocated altruism to be preferentially directed towards kin (Hamilton in J Theor Biol 7:1–16, 1964). We explored the possibility of kin recognition in the primitively eusocial halictid bee Lasioglossum malachurum by investigating the reliability of worker odour cues that can be perceived by workers to act as indicators of either nest membership or kinship. Cuticular and Dufour’s gland compounds varied significantly among colonies of L. malachurum, providing the potential for nestmate discrimination. A significant, though weak, negative correlation between chemical distance and genetic relatedness (r = −0.055, p < 0.001) suggests a genetic component to variation in cuticular bouquet, but odour cues were not informative enough to discriminate between different degrees of relatedness within nests. This pattern of variation was similar for Dufour’s gland bouquets. The presence of unrelated individuals within nests that are not chemically different from their nestmates suggests that the discrimination system of L. malachurum is prone to acceptance errors. Compounds produced by colony members are likely combined to generate a gestalt colony chemical signature such that all nestmates have a similar smell. The correlation between odour cues and nest membership was greater for perceived compounds than for non-perceived compounds, suggesting that variability in perceived compounds is a result of positive selection for nestmate recognition despite potentially stabilising selection to reduce variability in odour differences and thereby to reduce costs derived from excessive intracolony nepotistic behaviour.     

Nest separation and the dynamics of the Gestalt odor in the polydomous ant Cataglyphis iberica (Hymenoptera, Formicidae)

In the polydomous ant species Cataglyphis iberica, nests belonging to the same colony are completely separated during hibernation. In order to examine whether this separation induces changes both in the hydrocarbon profile and in recognition ability between adult nestmates, we separated groups of workers for several months under two different conditions: at hibernation temperature and at room temperature. At room temperature, recognition remained unchanged but separation led to longer mutual antennations relative to non-separated controls. When half of a colony was placed under hibernation conditions, antennal interactions also increased in duration and a few aggressive interactions emerged between separated ants. This aggressiveness never reached the intercolonial level observed in this species. In both cases, the hydrocarbon profiles showed differences between individuals after separation while remaining homogeneous within each nest. This chemical modification may induce the longer antennations observed. After separated groups were reunited, individuals recovered their previous antennation pattern and a convergence in hydrocarbon profiles was again observed. These concurrent observations suggest that hydrocarbons are transferred between nestmates. In C. iberica, the formation of the colonial odor seems to follow the “Gestalt” model which allows all satellite nests of a colony to have a common colonial odor. In the field, temporary nest isolation during hibernation may induce divergence between satellites. The role of adult transport in connecting nests during the active season to obtain an efficient Gestalt odor is discussed. Received: 16 June 1997 / Accepted after revision: 25 October 1997     

Nestmate recognition in three species of stenogastrine wasps (Hymenoptera,Vespidae)

The capacity to recognise a conspecific intruder was investigated in Parischnogaster jacobsoni, Liostenogaster flavolineata and L. vechti, three species of primitively social wasps of the subfamily Stenogastrinae. Results of behavioural experiments carried out in the field showed that females of all three species react pacifically if presented with female nestmates, but aggressively reject an intruder from a conspecific colony. As L. flavolineata and L. vechti both build large clusters of nests, often very close to each other, the recognition capacity among females from different nests, but in the same conspecific cluster, was also investigated. Females of both species were more aggressive towards females from a different colony in the same cluster than towards their female nestmates. Additional experiments on L. flavolineata showed that there was no difference in reaction towards females from colonies nearer or further from the tested colony but within the same cluster, nor towards females from a different cluster. The capacity to recognise an alien conspecific nest containing immature brood was investigated in P. jacobsoni. Adult females of this species, invited to land on an alien nest which had experimentally been exchanged for their own, accepted the new nest and partially destroyed the immature brood. The behaviour of the females when they land on an alien nest, however, suggests that they do recognise the nest as foreign. Acceptance of foreign nests coupled with low immature brood destruction is probably due to the high energetic costs of egg-deposition and larval rearing in stenogastrine wasps. These results suggest that nestmate recognition in these wasps is very efficient, even though they belong to the most primitive subfamily of social wasps. Received: 16 April 1996/Accepted after revision: 9 August 1996     

Nestmate discrimination in social wasps (Polistes metricus,Hymenoptera: Vespidae)

Summary Laboratory-overwintered paper wasp (Polistes metricus) females preferentially associate on new nests with former nestmates (presumably their sisters) rather than with non-nestmates in the absence of cues associated with their natal nests or nest sites. Females isolated from conspecifics and nests for 74–99 days retain the ability to discriminate nestmates from nonnestmates.     

Mating system of the cooperatively breeding noisy miner Manorina melanocephala,as revealed by DNA profiling

In this study we examined parentage within broods of the cooperatively breeding noisy miner using multi-locus DNA profiling. Previous studies of noisy miners described them as highly promiscuous, leading to the suggestion that promiscuous mating behaviour was a tactic used by females to recruit males as provisioners to their nests (Dow 1978). At our study site in south-east Queensland, Australia, we found that both multiple and extra-group paternity (i.e. a female mating with a male outside the group of male provisioners at her nest) were rare. In nests where multiple paternity was possible (i.e. clutch size > 1) 97% of 31 broods were sired by only a single male. Overall, 96.5% of all nestlings (n = 85) were the result of monogamous matings. Also, at the vast majority of nests, the male that sired the nestlings was also the main provisioner among all male nest attendants. Our results show that the mating system of the noisy miner can no longer be considered cooperative polyandry (Brown 1987) but is clearly genetic monogamy. We discuss the implications of this finding for understanding the complex social system of this species.     

Extended family structure in the ant Formica paralugubris: the role of the breeding system

In populations of various ant species, many queens reproduce in the same nest (polygyny), and colony boundaries appear to be absent with individuals able to move freely between nests (unicoloniality). Such societies depart strongly from a simple family structure and pose a potential challenge to kin selection theory, because high queen number coupled with unrestricted gene flow among nests should result in levels of relatedness among nestmates close to zero. This study investigated the breeding system and genetic structure of a highly polygynous and largely unicolonial population of the wood ant Formica paralugubris. A microsatellite analysis revealed that nestmate workers, reproductive queens and reproductive males (the queens' mates) are all equally related to each other, with relatedness estimates centring around 0.14. This suggests that most of the queens and males reproducing in the study population had mated within or close to their natal nest, and that the queens did not disperse far after mating. We developed a theoretical model to investigate how the breeding system affects the relatedness structure of polygynous colonies. By combining the model and our empirical data, it was estimated that about 99.8% of the reproducing queens and males originated from within the nest, or from a nearby nest. This high rate of local mating and the rarity of long-distance dispersal maintain significant relatedness among nestmates, and contrast with the common view that unicoloniality is coupled with unrestricted gene flow among nests. Received: 8 February 1999 / Received in revised form: 15 June 1999 / Accepted: 19 June 1999     

Relatedness among nestmates in a primitively social wasp,Cerceris antipodes (Hymenoptera: Sphecidae)

Summary Kin selection, acting through high levels of relatedness, may be an important promoter in the evolution of nest sharing. Cerceris antipodes is a sphecid wasp that shares nests in contrast to the majority of sphecids where only one female occupies a nest. Nest sharing results from females remaining in their natal nests and females moving to already occupied nests. Average relatedness among nestmates of C. antipodes was calculated from allele frequencies of phosphoglucomutase to determine whether nests were usually shared by close relatives. Relatedness among nestmates was high (0.5 to 0.6) at one aggregation in two consecutive years. Preferential association of relatives away from the natal nest can be inferred from these high values combined with the frequency of nest switching observed. Estimates of relatedness were lower (about 0.3) and associated with large standard errors at 3 other aggregations. Inbreeding and relatedness between neighbouring nests were only significant at the aggregation with the fewest females. This may be a result of the small effective population size. The levels of relatedness observed are consistent with kin selection through relatedness being an important factor in the evolution of nest sharing.     

Nestmate recognition and the genetic relatedness of nests in the ant Formica pratensis

Genetic relatedness of the mound-building ant Formica pratensis was determined by means of microsatellite DNA polymorphism, and its impact on nestmate recognition was tested in a population in Southern Sweden (Oeland). Recognition between nests was measured by testing aggression levels between single pairs of workers. The genetic distances of nests (Nei's genetic distance) and the spatial distance of nests were correlated and both showed a strong relation to the aggression behavior. Multiple regression analysis revealed a stronger impact of genetic relatedness rather than spatial distances on aggression behavior. Neighbouring nests were more closely related than distant nests, which may reflect budding as a possible spreading mechanism. The genetic distance data showed that nestmate recognition was strongly genetically influenced in F. pratensis. Received: 2 October 1997 / Accepted after revision: 10 January 1998     

Intranest relatedness and nestmate recognition in the meadow ant Formica pratensis (R.)

The impact of intranest relatedness on nestmate recognition was tested in a population of polydomous and monodomous nests of the mound-building ant Formica pratensis. Nestmate recognition was evaluated by testing aggression levels between 37 pairs of nests (n=206 tests). Workers from donor colonies were placed on the mounds of recipient nests to score aggressive interactions among workers. A total of 555 workers from 27 nests were genotyped using four DNA microsatellites. The genetic and spatial distances of nests were positively correlated, indicating budding and/or fissioning as spread mechanisms. Monodomous and polydomous nests did not show different aggression levels. Aggression behavior between nests was positively correlated with both spatial distance and intranest relatedness of recipient colonies, but not with genetic distance or intranest relatedness of donor colonies. Multiple regression analysis revealed a stronger effect of spatial distance than of genetics on aggression behavior in this study, indicating that the relative importance of environment and genetics can be variable in F. pratensis. Nevertheless, the positive regression between intranest relatedness of recipient colonies and aggression in the multiple analysis supports earlier results that nestmate recognition is genetically influenced in F. pratensis and further indicates that foreign label rejection most likely explains our data.     

Relatedness,recognition errors,and colony fusion in the termite <Emphasis Type="Italic">Nasutitermes corniger</Emphasis>

Loss of aggression between social groups can have far-reaching effects on the structure of societies and populations. We tested whether variation in the genetic structure of colonies of the termite Nasutitermes corniger affects the probability of aggression toward non-nestmates and the ability of unrelated colonies to fuse. We determined the genotypes of workers and soldiers from 120 colonies at seven polymorphic microsatellite loci. Twenty-seven colonies contained offspring of multiple founding queens or kings, yielding an average within-colony relatedness of 0.33. Genotypes in the remaining 93 colonies were consistent with reproduction by a single queen and king or their progeny, with an average within-colony relatedness of 0.51. In standardized assays, the probability of aggression between workers and soldiers from different colonies was an increasing function of within-colony relatedness. The probability of aggression was not affected significantly by the degree of relatedness between colonies, which was near zero in all cases, or by whether the colonies were neighbors. To test whether these assays of aggression predict the potential for colony fusion in the field, we transplanted selected nests to new locations. Workers and soldiers from colonies that were mutually tolerant in laboratory assays joined their nests without fighting, but workers and soldiers that were mutually aggressive in the assays initiated massive battles. These results suggest that the presence of multiple unrelated queens or kings promotes recognition errors, which can lead to the formation of more complex colony structures.     

Odour transfer in stingless bee marmelada (Frieseomelitta varia) demonstrates that entrance guards use an “undesirable–absent” recognition system

In group-level recognition, discriminators use sensory information to distinguish group members and non-members. For example, entrance guards in eusocial insect colonies discriminate nestmates from intruders by comparing their odour with a template of the colony odour. Despite being a species-rich group of eusocial bees closely related to the honey bees, stingless bee nestmate recognition is a relatively little-studied area. We studied Frieseomelitta varia, a common Brazilian species of stingless bee known as marmelada. By measuring the rejection rates of nestmate and non-nestmate worker bees by guards, we were able to show that guards became significantly less accepting (from 91 to 46%) of nestmates that had acquired odour cues from non-nestmate workers; however, guards did not become significantly more accepting (from 31 to 42%) of non-nestmates that had acquired equivalent amounts of odour cues from the guard’s nestmates. These data strongly suggest that guards use an “undesirable–absent” system in recognition, whereby incoming conspecific workers are only accepted if undesirable cues are absent, despite the presence of desirable cues. We suggest that an undesirable–absent system is adaptive because robbing by conspecifics may be an important selective factor in F. varia, which would lead to selection for a non-permissive acceptance strategy by guards.     

Variation in colony structure in the subterranean termite Reticulitermes flavipes

The genetic organization of colonies of the subterranean termite Reticulitermes flavipes in two subpopulations in Massachusetts was explored using five polymorphic allozymes and double-strand conformation polymorphism (DSCP) analysis of the mitochondrial control region. Empirically obtained estimates of worker relatedness and F-statistics were compared with values generated by computer simulations of breeding schemes to make inferences about colony organization. In one study site (G), worker genotypes indicated the presence of a mixture of colonies headed by monogamous outbred primary reproductives and colonies headed by inbreeding neotenic reproductives, both colony types having limited spatial ranges. A second site (S) was dominated by several large colonies with low relatedness among nestmates. Mixed DSCP haplotypes in three colonies indicated that nestmates had descended from two or three unrelated female reproductives. Computer simulations of breeding schemes suggested that positive colony inbreeding coefficients at site S resulted from either commingling of workers from different nests or different colonies. Such an exchange of workers between nests corresponds to the multiple-site nesting lifetype of many subterranean termites and resembles colony structure in polycalic Formica ants. Our study demonstrates considerable variation in R. flavipes colony structure over a small spatial scale, including colonies headed by monogamous outbred primary reproductives, colonies containing multiple inbred neotenic reproductives and large polydomous colonies containing the progeny of two or more unrelated queens, and suggests that the number of reproductives and nestmate relatedness change with colony age and size.     

The kin recognition system of carpenter ants (Camponotus spp.)

Summary Hamilton's (1964) hypothesis linking haplodiploidy and eusociality in the Hymenoptera could be reconciled with the occurrence of polygyny and multiple insemination if workers are able to distinguish full (3/4 related) sisters from other familiar matri- and patrilines within the colony, and direct altruistic behavior toward them preferentially. We examined this possibility in small genetically mixed nests of the carpenter ant Camponotus floridanus, formed by the transfer of worker pupae from two unrelated source colonies. In 120 h of observation on 12 queenright and 12 queenless nests, more than 15,000 behavioral interactions were recorded. Workers antennated familiar nonkin significantly more frequently than familiar sisters. However, they failed to discriminate consistently between kin and non-kin in food exchanges and grooming. Aggressive behavior was occasionally observed in some queenless nests, but almost never in the presence of a queen. When aggression did occur, it was directed significantly more often toward non-kin. Though related adult workers did not cooperate preferentially, the biases in antennation and aggression do indicate an ability to discriminate familiar kin from familiar nonkin, which may be employed in other contexts such as the rearing of reproductive brood.     

Hierarchical perception of fertility signals and nestmate recognition cues in two dolichoderine ants

In social insects, queens are likely to “honestly” inform their nestmates of their presence and fertility status through pheromonal communication. Cuticular hydrocarbons (CHCs) have been reported to be effective nestmate discriminators and strongly suspected to act as fertility signals, at least in some species. The use of the same chemical bouquet (i.e., the CHC profile) to convey two fundamentally different information seems puzzling. However, a recent threshold model proposes a hierarchy in the discriminating processes, i.e., fertility signals can only be perceived if nestmate recognition has been reached (Le Conte and Hefetz, Annu Rev Entomol 53:523–542, 2008). Here, we developed a simple behavioral bioassay based on chemical recruitment toward a queen placed outside the nest in two dolichoderine ants (Linepithema humile and Tapinoma erraticum), which allowed us to investigate the interplay between fertility signaling and colonial recognition. Using queen corpses of various origins (nestmates or aliens) and physiological states (fertile or infertile; mated or unmated), we demonstrated that nestmate recognition cues clearly override fertility signals under our experimental conditions. Indeed, while nestmate infertile queens were largely ignored by the workers, nestmate fertile queens (mated or not) induced worker recruitment, whereas alien fertile queens did not and were aggressed by the workers.     

Seasonal nestmate recognition in the ant Formica exsecta

Under favorable conditions, the mound-building ant Formica exsecta may form polydomous colonies and can establish large nest aggregations. The lack of worker aggression towards nonnestmate conspecifics is a typical behavioral feature in such social organization, allowing for a free flux of individuals among nests. However, this mutual worker toleration may vary over the seasons and on spatial scales. We studied spatio-temporal variation of worker–worker aggression within and among nests of a polydomous F. exsecta population. In addition, we determined inter- and intracolony genetic relatedness by microsatellite DNA genotyping and assessed its effect on nestmate recognition. We found significant differences in the frequency of worker exchange among nests between spring, summer, and autumn. Moreover, we found significant seasonal variation in the level of aggression among workers of different nests. Aggression levels significantly correlated with spatial distance between nests in spring, but neither in summer nor in autumn. Multiple regression analysis revealed a stronger effect of spatial distances rather than genetic relatedness on aggressive behavior. Because nestmate discrimination disappeared over the season, the higher aggression in spring is most plausibly explained by cue intermixing during hibernation.     

The role of wax and resin in the nestmate recognition system of a stingless bee, Tetragonisca angustula

Recent research has shown that entrance guards of the stingless bee Tetragonisca angustula make less errors in distinguishing nestmates from non-nestmates than all other bee species studied to date, but how they achieve this is unknown. We performed four experiments to investigate nestmate recognition by entrance guards in T. angustula. We first investigated the effect of colony odours on acceptance. Nestmates that acquired odour from non-nestmate workers were 63% more likely to be rejected while the acceptance rate of non-nestmates treated with nestmate odour increased by only 7%. We further hypothesised that guards standing on the wax entrance tube might use the tube as an odour referent. However, our findings showed that there was no difference in the acceptance of non-nestmates by guards standing on their own colony’s entrance tube versus the non-nestmate’s entrance tube. Moreover, treatment of bees with nestmate and non-nestmate resin or wax had a negative effect on acceptance rates of up to 65%, regardless of the origin of the wax or resin. The role of resin as a source of recognition cues was further investigated by unidirectionally transferring resin stores between colonies. Acceptance rates of nestmates declined by 37% for hives that donated resin, contrasting with resin donor hives where acceptance of non-nestmates increased by 21%. Overall, our results confirm the accuracy of nestmate recognition in T. angustula and reject the hypothesis that this high level of accuracy is due to the use of the wax entrance tubes as a referent for colony odour. Our findings also suggest that odours directly acquired from resin serve no primary function as nestmate recognition cues. The lack of consistency among colonies plus the complex results of the third and fourth experiments highlight the need for further research on the role of nest materials and cuticular profiles in understanding nestmate recognition in T. angustula.     

Sociogenetic organization of the red ant Myrmica rubra

Knowledge of the sociogenetic organization determining the kin structure of social insect colonies is the basis for understanding the evolution of insect sociality. Kin structure is determined by the number and relatedness of queens and males reproducing in the colonies, and partitioning of reproduction among them. This study shows extreme flexibility in these traits in the facultatively polygynous red ant Myrmica rubra. Relatedness among worker nestmates varied from 0 to 0.82. The most important reason for this variation was the extensive variation in the queen number among populations. Most populations were moderately or highly polygynous resulting in low relatedness among worker nestmates, but effectively monogynous populations were also found. Polygynous populations also often tend to be polydomous, which is another reason for low relatedness. Coexisting queens were positively related in two populations out of five and relatedness was usually similar among workers in the same colonies. Due to the polydomous colony organization and short life span of queens, it was not possible to conclusively determine the importance of unequal reproduction among coexisting queens, but it did not seem to be important in determining the relatedness among worker nestmates. The estimates of the mating frequency by queens remained ambiguous, which may be due to variation among populations. In some populations relatedness among worker nestmates was high, suggesting monogyny and single mating by queens, but in single-queen laboratory nests relatedness among the worker offspring was lower, suggesting that multiple mating was common. The data on males were sparse, but indicated sperm precedence and no relatedness among males breeding in the same colony. A comparison of social organizations and habitat requirements of M. rubra and closely related M. ruginodis suggested that habitat longevity and patchiness may be important ecological factors promoting polygyny in Myrmica. Received: 15 May 1995/Accepted after revision: 17 October 1995     

Begging behavior and food acquisition by brown-headed cowbird nestlings

Understanding the selective forces that limit the exaggeration of begging signals is a critical issue in understanding the evolution of begging behavior. I studied the begging behavior of nestlings of the brown-headed cowbird (Molothrus ater), a brood parasite. In the nests of indigo buntings (Passerina cyanea), brown-headed cowbird nestlings received approximately twice as much food per hour than their host nestmates. I tested three hypotheses for the mechanism by which cowbirds acquired more food than their bunting nestmates: the size advantage hypothesis, the signal exaggeration hypothesis, and the novel begging behavior hypothesis. I found support for the hypotheses that cowbirds acquire more food as a result of their larger body size, and due to the exaggeration of begging signals that are not dependent on body size. I did not find support for the role of novel begging behaviors in cowbird food acquisition. These results suggest that food acquisition by host chicks in unparasitized nests could be increased by the exaggeration of begging signals. Recent work suggests that such exaggeration may be limited by the risk of nest predation, but further studies are needed. Received: 12 December 1997 / Accepted after revision: 29 December 1997     

The effect of colony size and starvation on food flow in the fire ant,Solenopsis invicta (Hymenoptera: Formicidae)

Summary Food-sharing experiments were performed with laboratory colonies of Solenopsis invicta containing 1000, 10,000, or 20,000 workers and starved for 0, 3, 7, or 14 days. The effect of these variables was measured on the uptake of radioactive sugar water (1 M) by 1% of the colony's workers and on the trophallactic flow of food from these foragers to the remainder of the colony.Patterns of food distribution in small colonies differed significantly from those in larger nests. In 1000-ant nests, small workers more frequently received food than large workers, but in bigger colonies the opposite occurred.Fire ants were adept at distributing sugar water, with food from a few workers rapidly reaching the majority of the colony as foragers donate their crop contents to groups of recipients and these recipients may themselves act as donors.Foragers respond to colony starvation by individually taking up more food and sharing this fluid with a greater proportion of nestmates. Even foragers from satiated colonies can retrieve at least small amounts of liquid.The forager's state of hunger plays an important role in regulating food distribution. In sugar-satiated nests, previously starved foragers are highly successful at passing on labelled sugar whereas prviously fed foragers are not.     

Queuing for dominance: gerontocracy and queue-jumping in the hover wasp <Emphasis Type="Italic">Liostenogaster flavolineata</Emphasis>

The mechanisms through which dominance is inherited within social groups vary from direct interactions such as fighting to non-confrontational conventions. Liostenogaster flavolineata is a primitively eusocial hover wasp in which one female, the ‘dominant’, is the only reproductive upon the nest. The remaining females, although capable of reproduction, behave as helpers. In this study, we investigate the rules by which helpers inherit dominance. We removed successive dominants from 56 nests and recorded accession on un-manipulated nests. The results showed that L. flavolineata has a strict age-based inheritance queue: new dominants are the oldest female in their groups 87% of the time. Thirteen cases of queue-jumping were found in which young individuals were able to supplant older nestmates and inherit dominance precociously. Queue jumpers did not differ from other wasps in terms of relatedness to other group members or body size. Individuals that had previously worked less hard than other females of equivalent rank were significantly more likely to later jump the queue. Queue-jumping may represent a cheating strategy or could indicate that the rule for inheriting dominance is not based purely on relative age. We also discuss possible reasons why age-based queuing has evolved and its potential to promote the evolution of helping behaviour.