Cuticular Chemical Profiles Research Articles

Genetic distance and age affect the cuticular chemical profiles of the clonal ant Cerapachys biroi.

Although cuticular hydrocarbons (CHCs) have received much attention from biologists because of their important role in insect communication, few studies have addressed the chemical ecology of clonal species of eusocial insects. In this study we investigated whether and how differences in CHCs relate to the genetics and reproductive dynamics of the parthenogenetic ant Cerapachys biroi. We collected individuals of different ages and subcastes from several colonies belonging to four clonal lineages, and analyzed their cuticular chemical signature. CHCs varied according to colonies and clonal lineages in two independent data sets, and correlations were found between genetic and chemical distances between colonies. This supports the results of previous research showing that C. biroi workers discriminate between nestmates and non-nestmates, especially when they belong to different clonal lineages. In C. biroi, the production of individuals of a morphological subcaste specialized in reproduction is inversely proportional to colony-level fertility. As chemical signatures usually correlate with fertility and reproductive activity in social Hymenoptera, we asked whether CHCs could function as fertility-signaling primer pheromones determining larval subcaste fate in C. biroi. Interestingly, and contrary to findings for several other ant species, fertility and reproductive activity showed no correlation with chemical signatures, suggesting the absence of fertility related CHCs. This implies that other cues are responsible for subcaste differentiation in this species.

Polydomy in the antEctatomma opaciventre

Tropical ants commonly exhibit a hyper-dispersed pattern of spatial distribution of nests. In polydomous species, nests may be satellites, that is, secondary structures of the main nest, where the queen is found. In order to evaluate whether the ant Ectatomma opaciventre Roger (Formicidae: Ectatomminae) uses the strategy of building polydomous nests, the spatial distribution pattern of 33 nests in a 1,800 m2 degraded area located in Rio Claro, SP, Brazil, were investigated using the nearest neighbor method. To complement the results of this investigation, the cuticular chemical profile of eight colonies was analyzed using Fourier transform infrared photoacoustic spectroscopy (FTIR-PAS). The nests of E. opaciventre presented a hyper-dispersed or regular distribution, which is the most common in ants. The analysis of the cuticular hydrocarbons apparently confirmed the hypothesis that this species is polydomous, since the chemical profiles of all studied colonies with nests at different sites were very similar to the chemical signature of the single found queen and were also different from those of colonies used as control.

Polydomy in the ant Ectatomma opaciventre.

Tropical ants commonly exhibit a hyper-dispersed pattern of spatial distribution of nests. In polydomous species, nests may be satellites, that is, secondary structures of the main nest, where the queen is found. In order to evaluate whether the ant Ectatomma opaciventre Roger (Formicidae: Ectatomminae) uses the strategy of building polydomous nests, the spatial distribution pattern of 33 nests in a 1,800 m 2 degraded area located in Rio Claro, SP, Brazil, were investigated using the nearest neighbor method. To complement the results of this investigation, the cuticular chemical profile of eight colonies was analyzed using Fourier transform infrared photoacoustic spectrosco-py (FTIR-PAS). The nests of E. opaciventre presented a hyper-dispersed or regular distribution, which is the most common in ants. The analysis of the cuticular hydrocarbons apparently con-firmed the hypothesis that this species is polydomous, since the chemical profiles of all studied colonies with nests at different sites were very similar to the chemical signature of the single found queen and were also different from those of colonies used as control.

Major changes in the sex differences in cuticular chemical profiles of the western conifer seed bug (Leptoglossus occidentalis) after laboratory rearing

Chemical compounds covering the insect cuticle have several functions ranging from protection against water loss to inter- and intra-specific communication. Their composition is determined by several intrinsic and extrinsic factors. Among these factors, laboratory rearing has been poorly investigated even though it has a strong potential for biasing behavioral experiments. We selected an invasive species with unknown cuticular mixtures as a model. Our aim was to describe its mixtures and to determine if highly simplified laboratory rearing conditions interact with sexual signatures. We analyzed the cuticle by means of two different techniques – gas chromatography/mass spectrometry (GC/MS) and matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) – to obtain data on a wide range of compounds with different molecular weight. We found that both sets of chemicals showed correlations with gender but also that cuticular waxes detected by GC/MS were highly dependent on rearing conditions, with a strong bias in sexual dimorphism. Conversely, the heavier signatures detected by MALDI-TOF showed a less clear diversification between sexes, although the discrimination power was unaffected by rearing conditions. The biological and practical implications of our findings are discussed.

A conserved fertility signal despite population variation in the cuticular chemical phenotype of the trap-jaw antOdontomachus brunneus

Contact pheromones in the form of cuticular hydrocarbons are widespread among insects. Eusocial insects present a special challenge for understanding the evolution of the cuticular hydrocarbon profile because this blend is responsible for multiple distinct roles such as nestmate recognition and signalling fertility status. This study investigates these two signalling roles of the hydrocarbon profile in the trap-jaw ant Odontomachus brunneus. We demonstrate that the cuticular hydrocarbon profile is highly variable across populations and provide evidence that these differences are used for nestmate discrimination. Through manipulative experiments we also show that (Z)-9-nonacosene (Z9:C29) is used as a fertility signal and its role is conserved across populations. Our data demonstrate that both fertility and nestmate signalling influence the cuticular hydrocarbon profile and specifically the relative abundance of Z9:C29 on the cuticle of O. brunneus. Our study suggests that natural selection works on the cuticular chemical profile through multiple regulatory pathways, diversifying nestmate signals while conserving fertility signals.

Specific recognition of reproductive parasite workers by nest-entrance guards in the bumble bee Bombus terrestris

BackgroundThe 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.ResultsChemical 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.ConclusionOur 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.

Non-destructive species identification of Drosophila obscura and D. subobscura (Diptera) using near-infrared spectroscopy

The vinegar flies Drosophila subobscura and D. obscura frequently serve as study organisms for evolutionary biology. Their high morphological similarity renders traditional species determination difficult, especially when living specimens for setting up laboratory populations need to be identified. Here we test the usefulness of cuticular chemical profiles collected via the non-invasive method near-infrared spectroscopy for discriminating live individuals of the two species. We find a classification success for wild-caught specimens of 85%. The species specificity of the chemical profiles persists in laboratory offspring (87–92% success). Thus, we conclude that the cuticular chemistry is genetically determined, despite changes in the cuticular fingerprints, which we interpret as due to laboratory adaptation, genetic drift and/or diet changes. However, because of these changes, laboratory-reared specimens should not be used to predict the species-membership of wild-caught individuals, and vice versa. Finally, we demonstrate that by applying an appropriate cut-off value for interpreting the prediction values, the classification success can be immensely improved (to up to 99%), albeit at the cost of excluding a considerable portion of specimens from identification.

Cleptoparasites, social parasites and a common host: Chemical insignificance for visiting host nests, chemical mimicry for living in

Social insect colonies contain attractive resources for many organisms. Cleptoparasites sneak into their nests and steal food resources. Social parasites sneak into their social organisations and exploit them for reproduction. Both cleptoparasites and social parasites overcome the ability of social insects to detect intruders, which is mainly based on chemoreception. Here we compared the chemical strategies of social parasites and cleptoparasites that target the same host and analyse the implication of the results for the understanding of nestmate recognition mechanisms. The social parasitic wasp Polistes atrimandibularis (Hymenoptera: Vespidae), and the cleptoparasitic velvet ant Mutilla europaea (Hymenoptera: Mutillidae), both target the colonies of the paper wasp Polistes biglumis (Hymenoptera: Vespidae). There is no chemical mimicry with hosts in the cuticular chemical profiles of velvet ants and pre-invasion social parasites, but both have lower concentrations of recognition cues (chemical insignificance) and lower proportions of branched alkanes than their hosts. Additionally, they both have larger proportions of alkenes than their hosts. In contrast, post-invasion obligate social parasites have proportions of branched hydrocarbons as large as those of their hosts and their overall cuticular profiles resemble those of their hosts. These results suggest that the chemical strategies for evading host detection vary according to the lifestyles of the parasites. Cleptoparasites and pre-invasion social parasites that sneak into host colonies limit host overaggression by having few recognition cues, whereas post-invasion social parasites that sneak into their host social structure facilitate social integration by chemical mimicry with colony members.

Characterization of cuticular hydrocarbons of diploid and haploid males, workers and queens of the stingless bee Melipona quadrifasciata

Males, queens and workers of stingless bees show differences in external morphology, behaviour and roles within a colony. In addition, each individual has a cuticular chemical signature responsible for mutual communication that is essential for maintaining the integrity of the colony. In this paper we characterize the cuticular hydrocarbon composition of newly emerged diploid and haploid males, workers and virgin queens of Melipona quadrifasciata by gas chromatography-mass spectrometry (GC/MS) analysis. This is the first time that the cuticular profile of diploid males in a species of stingless bee has been characterized. We found differences in the cuticular hydrocarbon composition among males, workers and virgin queens, recording both qualitative and quantitative differences among individuals of different phenotypes. However, no compound was found exclusively in diploid males. The cuticular chemical profiles of haploid and diploid males were very similar to those of workers. Moreover, the cuticular lipids of males and workers were significantly different from those of queens. Tricosane, pentacosene-2 and 7-methyl-heptacosane were the compounds responsible for this significant separation. This result correlates with the behavioural and morphological differences among these phenotypes.

Cuticular Chemistry of Two Social Forms in a Facultatively Polygyne Ant (Hymenoptera: Formicidae:Formica truncorum)

In monogyne (single queen) ant colonies, worker aggression against intruders, including newly-mated sister queens (gynes) seeking readoption, prevents the development of multi-queen colonies. Some ant species, however, produce multiple queen colonies (polygyny) via the adoption of new gynes in response to ecological, genetic and social parameters. Cuticular chemistry mediates nestmate recognition as well as plays a role in intra-colonial regulation of reproduction. Although aggression against intruders is diminished in some polygyne species, apparently due to depauperate allelic diversity at loci that code for recognition cues or a loss of receptor sites, the mechanism underlying the development of polygyny is largely unknown. Here, we examine the cuticular chemical profiles of new gynes at several stages post-emergence, males and workers from monogyne and polygyne colonies in the facultatively polygyne ant Formica truncorum. Individuals from the two colony types were chemically distinguishable, as were gynes according to developmental stage post-emergence (callow versus mature) and with respect to wing presence and mating status. Overall profile complexity did not differ between populations. These results suggest an association of cuticular hydrocarbon profiles with dispersal tendencies and that the disparate tendency of workers from monogyne and polygyne colonies to adopt new gynes is due to a complex interaction between gyne cuticular chemistry and differential worker sensitivity to those cues.

Sick ants become unsociable

Parasites represent a severe threat to social insects, which form high-density colonies of related individuals, and selection should favour host traits that reduce infection risk. Here, using a carpenter ant (Camponotus aethiops) and a generalist insect pathogenic fungus (Metarhizium brunneum), we show that infected ants radically change their behaviour over time to reduce the risk of colony infection. Infected individuals (i) performed less social interactions than their uninfected counterparts, (ii) did not interact with brood anymore and (iii) spent most of their time outside the nest from day 3 post-infection until death. Furthermore, infected ants displayed an increased aggressiveness towards non-nestmates. Finally, infected ants did not alter their cuticular chemical profile, suggesting that infected individuals do not signal their physiological status to nestmates. Our results provide evidence for the evolution of unsociability following pathogen infection in a social animal and suggest an important role of inclusive fitness in driving such evolution.

Attracting Predators without Falling Prey: Chemical Camouflage Protects Honeydew‐Producing Treehoppers from Ant Predation

Predaceous ants are dominant organisms on foliage and represent a constant threat to herbivorous insects. The honeydew of sap-feeding hemipterans has been suggested to appease aggressive ants, which then begin tending activities. Here, we manipulated the cuticular chemical profiles of freeze-dried insect prey to show that chemical background matching with the host plant protects Guayaquila xiphias treehoppers against predaceous Camponotus crassus ants, regardless of honeydew supply. Ant predation is increased when treehoppers are transferred to a nonhost plant with which they have low chemical similarity. Palatable moth larvae manipulated to match the chemical background of Guayaquila's host plant attracted lower numbers of predatory ants than unchanged controls. Although aggressive tending ants can protect honeydew-producing hemipterans from natural enemies, they may prey on the trophobionts under shortage of alternative food resources. Thus chemical camouflage in G. xiphias allows the trophobiont to attract predaceous bodyguards at reduced risk of falling prey itself.

Social context predicts recognition systems in ant queens

Recognition of group-members is a key feature of sociality. Ants use chemical communication to discriminate nestmates from intruders, enhancing kin cooperation and preventing parasitism. The recognition code is embedded in their cuticular chemical profile, which typically varies between colonies. We predicted that ants might be capable of accurate recognition in unusual situations when few individuals interact repeatedly, as new colonies started by two to three queens. Individual recognition would be favoured by selection when queens establish dominance hierarchies, because repeated fights for dominance are costly; but it would not evolve in absence of hierarchies. We previously showed that Pachycondyla co-founding queens, which form dominance hierarchies, have accurate individual recognition based on chemical cues. Here, we used the ant Lasius niger to test the null hypothesis that individual recognition does not occur when co-founding queens do not establish dominance hierarchies. Indeed, L. niger queens show a similar level of aggression towards both co-foundresses and intruders, indicating that they are unable of individual recognition, contrary to Pachycondyla. Additionally, the variation in chemical profiles of Lasius and Pachycondyla queens is comparable, thus informational constraints are unlikely to apply. We conclude that selection pressure from the social context is of crucial significance for the sophistication of recognition systems.

Odour and Colour Similarity in Two Species of Gregarious Carabid Beetles (Coleoptera) from the Crati Valley, Southern Italy: A Case of Müllerian Mimicry?

Bombardier beetles of the genus Brachinus (Carabidae) are disagreeable prey because they discharge irritating quinones. Brachinus beetles live in aggregations and display warning colours. Like Brachinus beetles, Anchomenus dorsalis beetles produce methylsalicylate, and present a similar colour pattern. Anchomenus dorsalis beetles are usually found within Brachinus aggregations. Our aim was to investigate the similarity of cuticular chemical profiles of these two species to test the hypothe- sis of interspecific chemical mimicry. We investigated the cuticular composition of A. dorsalis, B. sclo- peta, and Poecilus cupreus. Poecilus cupreus, a non-aposematic carabid commonly found in Brachinus spp and Anchonemus dorsalis aggregations, was used as a control. The cuticular profiles of the three species include 48 different hydrocarbons. The cuticular hydrocarbon profiles of the three species of carabids were different, but individuals of Brachinus were chemically more similar to those of Ancho- menus than to those of Poecilus; in turn, individuals of Poecilus were more similar to Anchomenus than to Brachinus. We suggest that A. dorsalis is possibly mimicking the cuticular profile of B. sclopeta as an effective antipredator strategy. Brachinus sclopeta may benefit from a reduction of individual preda- tion risk due to an increased number of aggregated preys (dilution effect), while A. dorsalis may increase the benefit both from the dilution effect and from the greater chemical defense of B. sclopeta.

Food resources, chemical signaling, and nest mate recognition in the ant Formica aquilonia

Animals such as social insects that live in colonies can recognize intruders from other colonies of the same or different species using colony-specific odors. Such colony odors usually have both a genetic and an environmental origin. When within-colony relatedness is high (i.e., one or very few reproductive queens), colonies comprise genetically distinct entities, and recognition based on genetic cues is reliable. However, when nests contain multiple queens and colonies comprise multiple nests (polydomy), the use of purely genetically determined recognition labels may become impractical. This is due to high within-colony genetic heterogeneity and low between-colony genetic heterogeneity. This may favor the use of environmentally determined recognition labels. However, because nests within polydomous colonies may differ in their microenvironment, the use of environmental labels may also be impractical unless they are actively mixed among the nests. Using a laboratory experiment, we found that both isolation per se and diet composition influenced the cuticular chemical profiles in workers of Formica aquilonia. In addition, the level of aggression increased when both the proportions of dietary ingredients and the availability of food were altered. This suggests that increased aggression was mediated by changes in the chemical profile and that environmental cues can mediate recognition between colonies. These results also suggest that the underlying recognition cues are mutable in response to extrinsic factors such as the amount and the composition of food. Key words: aggressive behavior, colony odor, cuticular hydrocarbons, diet, social insects. [Behav Ecol]

Individual Recognition in Ant Queens

Personal relationships are the cornerstone of vertebrate societies, but insect societies are either too large for individual recognition, or their members were assumed to lack the necessary cognitive abilities . This paradigm has been challenged by the recent discovery that paper wasps recognize each other's unique facial color patterns . Individual recognition is advantageous when dominance hierarchies control the partitioning of work and reproduction . Here, we show that unrelated founding queens of the ant Pachycondyla villosa use chemical cues to recognize each other individually. Aggression was significantly lower in pairs of queens that had previously interacted than in pairs with similar social history but no experience with one another. Moreover, subordinates discriminated familiar and unfamiliar dominants in choice experiments in which physical contact, but not odor perception, was prevented and in tests with anaesthetized queens. The cuticular chemical profiles of queens were neither associated with dominance nor fertility and, therefore, do not represent status badges , and nestmate queens did not share a common odor. Personal recognition facilitates the maintenance of stable dominance hierarchies in these small societies. This suggests that the ability to discriminate between individual traits is selected for when it incurs net benefits for the resolution of conflict.

Dynamics of cuticular chemical profiles of Polistes dominulus workers in orphaned nests (Hymenoptera, Vespidae)

We analysed changes in cuticular hydrocarbon signatures of workers in orphaned colonies of the paper wasp Polistes dominulus. In natural conditions, workers and foundresses possess characteristic cuticular signatures, and foundresses are further distinguishable, both behaviourally and chemically, on the basis of their rank in a reproductive dominance hierarchy. In our study, several workers were found to develop their ovaries and produce cuticular signatures resembling those of dominant foundresses, while remaining workers possessed undeveloped ovaries and had cuticular blends characteristic of subordinate foundresses. Workers that did not develop their ovaries had changed epicuticular signatures, demonstrating that the mixture of hydrocarbons of worker individuals is strongly dependent on social role and environment. Our results suggest that the composition of epicuticular lipids is not determined at the pre-imaginal stage, and that physiological pathways leading to cuticular chemical changes are similar in foundresses and workers of P. dominulus.

Bumblebee inquilinism in Bombus�(Fernaldaepsithyrus) sylvestris (Hymenoptera, Apidae): behavioural and chemical analyses of host-parasite interactions

The bumblebee Bombus sylvestris is an obligate social inquiline of B. pratorum and nest- invading females are known to integrate themselves into host colonies, usually without any aggression. We investigated whether cuticular compounds could be involved in discrimination processes in common bumblebees, and whether they play a role in the social integration of inquilines in host nests. We tested nestmate recognition in B. terrestris workers, and analysed cuticular chemical profiles of hairs taken from allocolonial individuals. Bumblebees belonging to the same colony shared a common odour. Then, we investigated how B. sylvestris females integrated into colonies of natural and non-natural hosts, B. pratorum and B. terrestris, respectively. Inquiline females apparently succeeded in entering a host colony by expressing non-aggressive behaviours and had no chemical signature; subsequently, acquisition of a chemical signature similar to the host colony might facilitate their integration into the host nest. Bombus sylvestris / inquilinism / discrimination process / chemical signature / non-aggressive strategy

Hibernation clustering and its consequences for associative nest foundation in Polistes dominulus (Hymenoptera Vespidae)

Various studies indicate that cuticular hydrocarbons (CHCs) act as cues for nestmate recognition in paper wasps and other social insects. A recent study showed that associative nest foundation in Polistes dominulus is mainly performed by foundresses coming from the same locality. In the present study, we induced future foundresses of P. dominulus collected in two different localities to hibernate in the laboratory in aggregates of individuals from the same or different localities. After 2 months of hibernation, foundresses from different localities but from the same experimental cluster did not show any preference to associate, at the time of nest foundation, with individuals coming from the same original locality. The cuticular chemical profiles of individuals from the mixed hibernation clusters were quite similar and significantly different from those of individuals which hibernated with other foundresses from the same locality. These findings suggest that, in this species, mechanisms other than nestmate chemical recognition play a major role in the spring association of gynes during the foundation of a new nest.

Variation in cuticular hydrocarbon signatures, hormonal correlates and establishment of reproductive dominance in a polistine wasp

In many social insects the relationship between reproductive dominance and physiological correlates is poorly understood. Recent evidence now strongly suggests that cuticular hydrocarbons are important in reproductive differentiation in these societies where they are used as signals of ovarian activity in reproductive females. In this study we investigated the relationship between reproductive dominance, size of the corpora allata (CA, producer of Juvenile Hormone, JH) and the proportions of cuticular hydrocarbons present on the cuticle in overwintering foundresses and both associative (polygynous) and solitary (monogynous) pre-emergence colonies of the social wasp Polistes dominulus. Size of the CA was positively correlated with ovarian development in polygynous colonies. In contrast, solitary foundresses possessed significantly smaller CAs than dominant foundresses from polygynous nests, yet ovarian activity was similar for both female types. CA size variation was associated with variation in cuticular hydrocarbon proportions. Overwintering, solitary, dominant and subordinate (from associative nests) females all possessed distinctive cuticular chemical profiles revealed by multivariate discriminant analyses. Our data indicate that the social environment strongly affects reproductive physiology in this wasp, and we discuss the role of cuticular hydrocarbons in reproductive signaling in P. dominulus and other social insects.