Cuticular Profiles Research Articles

Influence of Host Plants and Tending Ants on the Cuticular Hydrocarbon Profile of a Generalist Myrmecophilous Caterpillar.

In myrmecophilous organisms, which live in symbiosis with ants, cuticular hydrocarbons (CHCs) play a pivotal role in interspecific communication and defense against chemical-oriented predators. Although these interactions form complex information webs, little is known about the influence of biotic environmental factors on the CHC profiles of myrmecophiles. Here, we analyzed the effect of different host plants and tending ants on the larval CHC profile of Synargis calyce (Lepidoptera: Riodinidae), a polyphagous species with facultative myrmecophily. Groups of caterpillars were fed individually with three host plant species (without tending ants), and with two tending ant species. Through gas chromatography analysis, we compared the cuticular profiles of treatments and found a high similarity between plants and caterpillars (65-82%), but a low similarity between caterpillars and their tending ants (30 - 25%). Cluster analysis showed that caterpillars, ants, and plants form distinct groups, indicating that S. calyce caterpillars have their own chemical profile. These results are similar to those observed for Lycaenidae caterpillars indicating that there is functional convergence in the chemical strategies used by myrmecophilous caterpillar species with similar ecology. Also, the results suggest that the cuticular compounds of S. calyce are primarily influenced by their host plants rather than their tending ants. Thus, we propose that these caterpillars present a trade-off between camouflage and directly informing their presence to ants, maintaining their unique chemical profile, though slightly affected by biotic environmental factors.

Cuticular lipid profiles of selected species of cyclocephaline beetles (Melolonthidae, Cyclocephalini).

Neotropical cyclocephaline beetles, a diverse group of flower-loving insects, significantly impact natural and agricultural ecosystems. In particular, the genus Cyclocephala, with over 350 species, displays polymorphism and cryptic complexes. Lacking a comprehensive DNA barcoding framework, accessible tools for species differentiation are needed for research in taxonomy, ecology, and crop management. Moreover, cuticular hydrocarbons are believed to be involved in sexual recognition mechanisms in these beetles. In the present study we examined the cuticular chemical profiles of six species from the genus Cyclocephala and two populations of Erioscelis emarginata and assessed their efficiency in population, species, and sex differentiation. Overall we identified 74 compounds in cuticular extracts of the selected taxa. Linear alkanes and unsaturated hydrocarbons were prominent, with ten compounds between them explaining 85.6% of species dissimilarity. Although the cuticular chemical profiles efficiently differentiated all investigated taxa, only C. ohausiana showed significant cuticular profile differences between sexes. Our analysis also revealed two E. emarginata clades within a larger group of 'Cyclocephala' species, but they were not aligned with the two studied populations. Our research underscores the significance of cuticular lipid profiles in distinguishing selected cyclocephaline beetle species and contemplates their potential impact as contact pheromones on sexual segregation and speciation.

Contact chemoreceptive mate recognition in Cerambyx welensii Küster (Coleoptera: Cerambycidae)

Abstract Cerambyx welensii Küster (Coleoptera: Cerambycidae) is a wood‐borer responsible for the decline of Mediterranean oaks in open woodlands. To establish that contact pheromones are involved in mate recognition of C. welensii, we extracted the cuticular hydrocarbons by solid phase microextraction at pre‐reproductive, beginning and ending of the reproductive period, and by solvent extraction of prothorax and elytra. The extracts were analysed by GC–MS under electron impact and chemical ionization conditions. Cuticular hydrocarbon profiles varied according to the reproductive period, although differences between sexes were not significant. Two compounds, 11/13‐methylheptacosane and 11‐methylnonacosane were more abundant in females at the beginning and ending of the reproductive periods. Compound 11/13‐methylheptacosane was also more representative in female prothorax than in males, and 2‐methyloctacosane was richer in male elytra than in females. We also studied the role of cuticular hydrocarbons in mate recognition in arena bioassays. Treatment of solvent‐washed dead females and glass dummies with one female equivalent (FE) of cuticular extract elicited mating responses in males, especially at the beginning of the reproductive period, with copulation attempts reaching 61.9% on solvent‐washed dead females and 23.8% on dummies. The successive treatment with synthetic compounds approaching a male cuticular profile inhibited male response. Our results confirm that contact pheromones mediate mate recognition in C. welensii. Knowledge of the precise role played by the major compounds 11/13‐methylheptacosane and 11‐methylnonacosane and other minor compounds representative in female prothorax may contribute to the development of novel management strategies against C. welensii.

The Cuticular Hydrocarbons of Dasineura Oleae Show Differences Between Sex, Adult Age and Mating Status

In insects, cuticular lipids prevent water loss and act as semiochemicals. Because of their ecological function, the profile change across the insects’ sex and development offers insight into insect biology and possible tools for pest management. Here, the first work on cecidomyiid cuticular extracts is proposed considering Dasineura oleae (Diptera: Cecidomyiidae) males and females at different adult ages (0–12 h, 12–24 h, 24–36 h) and distinct sexual conditions (virgin and mated). A set of 49 compounds were recorded (12 alkanes, 1 monomethyl alkane, 11 fatty acids, 4 esters, 1 aldehyde, 1 allylbenzene, 1 amine, 1 flavonoid, 1 ketone, 1 phenol, 1 steradiene, 1 sterol, 1 terpene, 1 triterpene and 11 unknown compounds), and 18 of them showed significant differences between groups. Among alkanes, hexacosane (nC26) exhibited a decreasing trend from the youngest to the oldest females, while pentacosane (nC25) and nonacosane (nC29) showed a decreasing trend from 0 to 12 h to 12–24 h virgin females. In addition, nonadecane (nC19) was significantly more abundant in the youngest males compared to older males and females. The alkanes nC25, nC26 and nC29 have been reported to be age-related also in other dipterans, while nC19 has been described as gender-specific chemical cue for platygastrid parasitoids. Further behavioural trials and analyses are required to assign the specific ecological roles to the characterized compounds. Our results may contribute to develop new low-impact control strategies relying on the manipulation of D. oleae’s chemical communication (e.g. disruption of mating or species recognition).Highlights• Cuticular hydrocarbons are often involved in dipteran intraspecific communication.• We explored the cuticular profile of D. oleae at different age, sex, mating condition.• Five alkanes and one mono-methyl alkane showed differences among groups.• Linoleic acid is the most abundant compound in virgins, absent in mated insects.• Eleven compounds disappear in mated insects, but were present in all virgins.Graphical

Variation in contact chemical cues may mediate differential predator response in the colour polymorphic tortoise beetle,Chelymorpha alternans

AbstractDifferential predation on species with intraspecific colour variation has been explored in various systems and is often implicated as the driving force behind colour polymorphism maintenance.Here, investigation done on whether predation contributes to the maintenance of extensive colour variation in the Neotropical tortoise beetle,Chelymorpha alternans(Chrysomelidae). Recorded predation rates on different colour pattern phenotypes by three common, generalist invertebrate predators, and identified potential chemical signals of unpalatability.Predaceious mantids (Orthoptera, Mantidae) consumed no beetles, regardless of phenotype, whereas the giant orb‐weaving spider (Trichonephila clavipes; Araneidae) consumed all three beetle phenotypes. The carton‐nest ant,Azteca chartifex(Formicidae), displayed differential predation; the rufipennis phenotype ofC. alternanswas sometimes consumed, the metallic phenotype was never consumed, and the veraguensis phenotype was consumed in the first three encounters and subsequently discarded, suggesting a learned avoidance behaviour.Using gas chromatography–mass spectrometry, it was determined that cuticular hydrocarbon profiles were similar between the metallic and militaris‐a phenotypes. The rufipennis phenotype showed pronounced differences and displayed the greatest among‐individual variation in elytral cuticular profiles.Between‐phenotype variation in chemical cues, and differences in how predators receive those cues, may mediate predator response, and play a role in maintaining colour variation in this species.

Small hive beetle, Aethina tumida (Coleoptera: Nitidulidae): chemical profile of the cuticle and possible chemical mimicry in a honeybee (Apis mellifera) pest

The small hive beetle, Aethina tumida (Coleoptera: Nitidulidae), is an economically important pest of the Western honeybee, Apis mellifera (Hymenoptera: Apidae). We investigated the effect of rearing environment on the cuticular chemical profile of adult A. tumida, using hexane to extract the hydrocarbons and other compounds from the cuticles of beetles. Beetles were collected from A. mellifera colonies in Australia as well as reared in single sex laboratory cultures on different diets. We investigated whether rearing environment (laboratory vs. field, different apiaries, access to mating partners, diet) had any effect on cuticular hydrocarbons. Coupled gas chromatography–mass spectrometry analyses of the extracts showed that rearing environment had significant qualitative and quantitative effects on the hydrocarbons detected. The data support the hypothesis that cuticular profiles of A. tumida are contingent on environment, partitioning on the basis of rearing diet and source hives. The finding has implications for the regulation of interactions between A. tumida and honeybees and improvements in targeting of management strategies.

Sympatric cleptobiotic stingless bees have species-specific cuticular profiles that resemble their hosts

Stingless bees are the largest group of eusocial pollinators with diverse natural histories, including obligate cleptobionts (genus Lestrimelitta) that completely abandoned flower visitation to rely on other stingless bees for food and nest materials. Species of Lestrimeliita are thought to specialize upon different host species, and deception through chemical similarity has been proposed as a mechanism to explain this phenomenon. In the Yucatan Peninsula of Mexico, Scaptotrigona pectoralis is a species chemically distinct from, and not preferred as a host by, locally widespread Lestrimeliita niitkib; witnessing attacks on S. pectoralis colonies offered the opportunity to test the sensory deception hypothesis to cletoparasitism. Analysis of cuticular profiles revealed that the Lestrimelitta attacking S. pectoralis differed significantly in odour bouquet to L. niitkib and, in contrast, it resembled that of S. pectoralis. Further analyses, including morphometrics, mtDNA barcoding, and the examination of taxonomic features, confirmed the existence of two sympatric Lestrimelitta species. The results give support to the hypothesis of chemical deception as a cleptobiotic strategy in Lestrimelitta sp. This is the first evidence that sympatric cleptobionts of the same genus select hosts in accordance with species-specific cuticular profiles, with possible consequences for ecological adaptation and the evolution of these remarkable organisms and the community of stingless bee hosts.

Deciphering the chemical phenotype in Atta laevigata (Smith, 1858) (Hymenoptera: Formicidae): A relationship between polymorphism and cuticular hydrocarbons

In the Attina subtribe the division of labor among workers is based on different tasks performed by morphological subcastes. Considering that cuticular chemical compounds play important roles as protection against water loss and mediates interactions between nestmates, and that studies on the cuticular chemical profile of ants of the genus Atta are still scarce, the aim of this study was to evaluate the relationship between morphological and chemical phenotypes in the subcastes of Atta laevigata (Smith, 1858) workers. The cuticular chemical composition of the different subcastes was assessed by Gas Chromatography coupled to Mass Spectrometry technique. The compounds found in the cuticle of the different subcastes had carbon chain size between 18 and 31 atoms, with higher content of linear alkanes in workers and soldiers and higher content of branched alkanes in gardeners and generalists, probably related to the type of tasks they perform in their colonies, since some workers perform more intra- or extranidal tasks in relation to others. The results show significant qualitative differences in the cuticular composition of the different subcastes, with a greater relationship between them according to the environmental restrictions that each subcaste is more subjected to due to the role played in the colony.

Neglected Very Long-Chain Hydrocarbons and the Incorporation of Body Surface Area Metrics Reveal Novel Perspectives for Cuticular Profile Analysis in Insects.

Simple SummaryThe waxy layer covering the surface of most terrestrial insects is mainly composed of non-polar lipids termed cuticular hydrocarbons (CHCs). These have a long research history as important dual traits for both desiccation prevention and chemical communication. We analyzed CHC profiles of seven species of the insect order Blattodea (termites and cockroaches) with the most commonly applied chromatographic method, gas-chromatography coupled with mass spectrometry (GC-MS), and the more novel approach of silver-assisted laser desorption/ionization mass spectrometry (Ag-LDI-MS). Comparing these two analytical methods, we demonstrated that the conventional GC-MS approach does not provide enough information on the entire CHC profile range in the tested species. Ag-LDI-MS was able to detect very long-chain CHCs ranging up to C58, which remained undetected when solely relying on standard GC-MS analysis. Additionally, we measured the body surface areas of each tested species applying 3D scanning technology to assess their respective CHC amounts per mm2. When adjusting for body surface areas, proportional CHC quantity distributions shifted considerably between our studied species, suggesting the importance of including this factor when conducting quantitative CHC comparisons, particularly in insects that vary substantially in body size.Most of our knowledge on insect cuticular hydrocarbons (CHCs) stems from analytical techniques based on gas-chromatography coupled with mass spectrometry (GC-MS). However, this method has its limits under standard conditions, particularly in detecting compounds beyond a chain length of around C40. Here, we compare the CHC chain length range detectable by GC-MS with the range assessed by silver-assisted laser desorption/ionization mass spectrometry (Ag-LDI-MS), a novel and rarely applied technique on insect CHCs, in seven species of the order Blattodea. For all tested species, we unveiled a considerable range of very long-chain CHCs up to C58, which are not detectable by standard GC-MS technology. This indicates that general studies on insect CHCs may frequently miss compounds in this range, and we encourage future studies to implement analytical techniques extending the conventionally accessed chain length range. Furthermore, we incorporate 3D scanned insect body surface areas as an additional factor for the comparative quantification of extracted CHC amounts between our study species. CHC quantity distributions differed considerably when adjusted for body surface areas as opposed to directly assessing extracted CHC amounts, suggesting that a more accurate evaluation of relative CHC quantities can be achieved by taking body surface areas into account.

Identification of Cuticular and Web Lipids of the Spider Argiope bruennichi

Emerging evidence shows that the cuticular and silk lipids of spiders are structurally more diverse than those of insects, although only a relatively low number of species have been investigated so far. As in insects, such lipids might play a role as signals in various contexts. The wasp spider Argiope bruennichi has probably the best investigated chemical communication system within spiders, including the known structure of the female sex pheromone. Recently we showed that kin-recognition in A. bruennichi could be mediated through the cuticular compounds consisting of hydrocarbons and, to a much larger proportion, of wax esters. By use of mass spectrometry and various derivatization methods, these were identified as esters of 2,4-dimethylalkanoic acids and 1-alkanols of varying chain lengths, such as tetradecyl 2,4-dimethylheptadecanoate. A representative enantioselective synthesis of this compound was performed which proved the identifications and allowed us to postulate that the natural enantiomer likely has the (2R,4R)-configuration. Chemical profiles of the silk and cuticular lipids of females were similar, while male cuticular profiles differed from those of females. Major components of the male cuticular lipids were tridecyl 2,4-dimethyl-C17-19 alkanoates, whereas those of females were slightly longer, comprising tridecyl 2,4-dimethyl-C19-21 alkanoates. In addition, minor female-specific 4-methylalkyl esters were detected.

Side effects of a fungus-based biopesticide on stingless bee guarding behaviour

Pathogenic fungi have been used worldwide to control crop pests and are assumed to pose negligible threats to the survival of pollinators. Although eusocial stingless bees provide essential pollination services and might be exposed to these biopesticides in tropical agroecosystems, there is a substantial knowledge gap regarding the side effects of fungal pathogens on behavioural traits that are crucial for colony functioning, such as guarding behaviour. Here, we evaluated the effect of Beauveria bassiana on the sophisticated kin recognition system of Tetragonisca angustula, a bee with morphologically specialized entrance guards. By combining behavioural assays and chemical analyses, we show that guards detect pathogen-exposed nestmates, preventing them from accessing nests. Furthermore, cuticular profiles of pathogen-exposed foragers contained significantly lower amounts of linear alkanes than the unexposed ones. Such chemical cues associated with fungal conidia may potentially trigger aggression towards pathogen-exposed bees, preventing pathogen spread into and among colonies. This is the first demonstration that this highly abundant native bee seems to respond in a much more adaptive way to a potentially infectious threat, outweighing the costs of losing foraging workforce when reducing the chances of fungal pathogen outbreaks within their colonies, than honeybees do.

Family-specific chemical profiles provide potential kin recognition cues in the sexually cannibalistic spider Argiope bruennichi.

Kin recognition, the ability to detect relatives, is important for cooperation, altruism and also inbreeding avoidance. A large body of research on kin recognition mechanisms exists for vertebrates and insects, while little is known for other arthropod taxa. In spiders, nepotism has been reported in social and solitary species. However, there are very few examples of kin discrimination in a mating context, one coming from the orb-weaver Argiope bruennichi. Owing to effective mating plugs and high rates of sexual cannibalism, both sexes of A. bruennichi are limited to a maximum of two copulations. Males surviving their first copulation can either re-mate with the current female (monopolizing paternity) or leave and search for another. Mating experiments have shown that males readily mate with sisters but are more likely to leave after one short copulation as compared with unrelated females, allowing them to search for another mate. Here, we ask whether the observed behaviour is based on chemical cues. We detected family-specific cuticular profiles that qualify as kin recognition cues. Moreover, correlations in the relative amounts of some of the detected substances between sexes within families indicate that kin recognition is likely based on subsets of cuticular substances, rather than entire profiles.

Similar Is Not the Same – Mate Recognition in a Parasitoid Wasp

Finding and recognizing a suitable mate is a key prerequisite to reproductive success. Insects often recognize prospective mates using chemical cues and signals. Among these, cuticular lipids commonly serve for mate recognition at close range. The lipid layer on the surface of insects is comprised predominantly of cuticular hydrocarbons (CHCs), though more polar compounds may also be present. While the composition of the cuticular profile is typically species specific, many species additionally show differentiation between the sexes by sex specific compounds and/or compound ratios. It is often assumed that a clear sexual dimorphism of cuticular lipid profiles is a prerequisite for a potential function as sex pheromones. Both, sex specific single compounds or the profiles as a whole have been shown to serve as sex pheromones in parasitoid wasps. Here, we studied parasitoid wasps of the speciesTachinaephagus zealandicus(Encyrtidae, Hymenoptera). Chemical analyses revealed that this species presents a case where males and females produce the same set of CHCs in similar relative amounts. To test whether these wasps nonetheless can use the cuticular lipids for close range mate recognition, we tested the reaction of males toward freeze-killed conspecifics. Males showed copulation behavior exclusively toward dead females, but not toward dead males. Dead females from which the cuticular lipids had been removed did not elicit copulation behavior by tested males. Reapplication of female whole body extracts restored bioactivity, and males reacted with copulation attempts as often as toward the freeze-killed females. Bioassays with lipid fractions revealed that only the CHC fraction was bioactive on its own. Here, again, males reacted to female, but not to male CHCs. Our results indicate that these wasps are capable of using CHCs for close range sex recognition despite the similarity of male and female profiles.

Cues of dominance hierarchy, fertility and nestmate recognition in the primitively eusocial wasp Mischocyttarus parallelogrammus (Vespidae: Polistinae: Mischocyttarini)

Chemical communication is pivotal for social insects to ensure proper functioning of their colonies. Cuticular hydrocarbons (CHCs) are the most well-known class of compounds used to regulate different types of behavioural interaction within a social context. Queens of highly eusocial insects rely on the use of chemical communication to keep their reproductive monopoly, whereas queens of primitively eusocial insects often exert physical control to maintain reproductive dominance. However, in the past years, ample evidence has demonstrated that primitively eusocial insects also use chemical compounds to communicate. Based on this evidence, we aimed to elucidate whether CHCs carry some information regarding female hierarchical position, ovary activation and nestmate recognition in the primitively eusocial wasp Mischocyttarus parallelogrammus. Additionally, females were classified by their ovary activation. Finally, the cuticular profiles of females originating from different nests were compared to check whether CHCs convey information about their nest of origin. Our results confirmed that the chemical composition of alpha and subordinate females differed significantly in post-worker emergence nests, but that alpha and beta females surprisingly were not chemically different from each other in either of the colony phases. Furthermore, females with activated ovaries expressed a chemical profile distinct from that of females with non-activated ovaries. Lastly, we showed that CHCs might convey information about nest origin, since females hailing from different nests showed distinct chemical profiles. Based on our results, we conclude that CHCs might play a critical role in the nest-functioning of M. parallelogrammus, since they mirror social status.

Strategies of the beetle Oochrotus unicolor (Tenebrionidae) thriving in the waste dumps of seed‐harvesting Messor ants (Formicidae)

1. A diverse group of arthropods have adapted to the niches found inside the nests of social insects. Studies mostly focused on very specialised parasites residing in the brood chambers. However, the biology and strategies of symbionts occupying other niches, such as waste dumps, are underexplored.2. Using a series of complementary experiments, this study demonstrated that the Mediterranean beetle Oochrotus unicolor has adapted to the waste dump niche found in the nests of Messor harvester ants.3. Laboratory experiments confirmed field observations that the beetle preferentially resided in the refuse pits. Next, it was shown that the beetles readily consumed seeds and flour, whereas other food sources were poorly accepted and ant brood was never even eaten. The beetles did not elicit a strong aggression response in Messor ants, and they could tolerate very high densities of workers without clear costs. The beetles modestly mimicked the nest recognition cues of their Messor host. This imperfect mimicry could promote the adoption of the beetle in the ant colony, in concert with mechanical defence generated by its tank‐like body. Isolation of the beetle from its host did not significantly affect the beetle's chemical cuticular profile nor did it provoke elevated ant aggression, indicating that the beetle does not acquire the chemicals passively from its host.4. This paper discusses the fact that waste dumps in social insect nests are hotspots for arthropod symbionts. It shows that symbionts in this niche may employ behavioural, trophic and chemical strategies that are different from those found in other niches of social insect nests.

Pre-existing differences in putative fertility signals give workers the upper hand in ant reproductive hierarchies

In social groups, competition often gives rise to conflicts, which are regulated through a variety of mechanisms. In several social insect species, the conflict for male production that takes place between workers after queen loss, is regulated through the establishment of a reproductive hierarchy. A recent study of Neoponera apicalis showed that workers differ in their fertility levels in the presence of the queen and proposed that such idiosyncratic differences might influence access to the top of the hierarchy after queen loss. In this study, we therefore sought to characterize the influence of the initial heterogeneity in ovarian development and its chemical and behavioural correlates on the establishment of reproductive hierarchies among orphaned workers, which can only produce males. We monitored the chemical profile before and after hierarchy establishment in four groups of orphaned workers of N. apicalis morph 6. The analysis of the cuticular profiles showed that tricosane (n-C23) was highly correlated with ovarian development and could consequently act as a fertility signal in this ant. The relative amount of tricosane on the cuticle, both before and after the establishment of the hierarchy, was also correlated with the rank achieved within the hierarchy and with the expression of agonistic behaviours. Thus, our study experimentally shows that idiosyncratic differences in a putative fertility signal (and therefore presumably in ovarian activity) between workers in the queen's presence reliably predict the outcome of reproductive conflict after queen loss. We propose that this signal (together with an increased agonistic motivation of the more fertile workers) could play a major role in the regulation of dominance/submission behaviours, enabling the most fertile individuals to rapidly access top ranks and monopolize reproduction, thereby maximizing the global reproductive success of all colony workers while minimizing the costs associated with the expression of agonistic behaviour.

Defective Pollen Wall 3 (DPW3), a novel alpha integrin-like protein, is required for pollen wall formation in rice.

In flowering plants, pollen wall is a specialized extracellular cell-wall matrix surrounding male gametophytes and acts as a natural protector of pollen grains against various environmental and biological stresses. The formation of pollen wall is a complex but well-regulated process, which involves the action of many different genes. However, the genetic and molecular mechanisms underlying this process remain largely unknown. In this study, we isolated and characterized a novel rice male sterile mutant, defective pollen wall3 (dpw3), which displays smaller and paler anthers with aborted pollen grains. DPW3 encodes a novel membrane-associated alpha integrin-like protein conserved in land plants. DPW3 is ubiquitously expressed in anther developmental stages and its protein is localized to the plasma membrane, endoplasmic reticulum (ER) and Golgi. Anthers of dpw3 plants exhibited unbalanced anther cuticular profile, abnormal Ubisch bodies, disrupted callose deposition, defective pollen wall formation such as abnormal microspore plasma membrane undulation and defective primexine formation, resulting in pollen abortion and complete male sterility. Our findings revealed a novel and vital role of alpha integrin-like proteins in plant male reproduction.

Dear Enemy Phenomenon in the Ant Ectatomma brunneum (Formicidae: Ectatomminae): Chemical Signals Mediate Intraspecific Agressive Interactions

The integrity of ant colonies depends exclusively on social relationships between their individuals, especially the ability of communication between group members, which is mainly mediated through chemical signals. Another important feature of ant behavior is territory defense, since they need to gather large amounts of food to feed their larvae, males and breeding females. Thereby, ants might display behavioral strategies to defend their territories from intruders. Here we investigated whether Ectatomma brunneum displays the Dear Enemy Phenomenon, what is the relationship between Cuticular Hydrocarbon composition and levels of aggression during their intraspecific interactions and which compounds and/or classes of compounds might be the most important to modulate the level of aggression. To test our hypothesis, we evaluated the levels of aggression through behavioral observations during interactions between 23 pairs of colonies nested in two distinct sites at varied distances. Then, we analyzed the cuticular chemical profile of the individuals involved in the interactions, and compared these results with the levels of aggression displayed between colonies tested. The results allow us to confirm our hypothesis that the DEP occurs in E. brunneum. The higher tolerance between closer colonies can be explained due to their kinship level in addition to sharing the same microhabitats. The results also showed there are significant differences in CHCs profiles, especially between colonies nested at relatively greater distances, and it is likely that differences in content of some branched alkanes are the most important to establish these differences and, therefore, the levels of aggression during the interactions.

Cuticular hydrocarbons as cues of sex and health condition in Polistes dominula wasps

In the paper wasp Polistes dominula, cuticular hydrocarbons play a critical role to acquire information regarding conspecific individuals. However, the relationship between cuticular hydrocarbons, health status, and male sexually selected traits is poorly investigated. In this study, we characterized the cuticular hydrocarbon profile of adult male and female wasps, infected or not by the strepsipteran endoparasite Xenos vesparum, to assess whether the chemical signature provides information about sex and health status (parasite infection). Moreover, we tested whether the chemical profile reflects male quality as measured via morphological and behavioural (sexually selected) traits at leks. Our results showed that males and females had similar total amount of CHCs, quantitatively different profiles and, to a lesser extent, sex-specific chemical compounds. Cuticular profiles were influenced by the strepsipteran infection, and the effect was stronger in females (the primary host) than in males, according to the physiological castration of female but not of male hosts. Regarding territorial and non-territorial males, no significant difference emerged in their chemical profiles. Furthermore, sex-dimorphic visual signals (size, shape, and asymmetry of abdominal yellow spots) were related to the behavioural displays of territorial males. We hypothesize that cuticular hydrocarbons are potential multi-role cues to assess sex and health status in male and female wasps, in synergy with visual signals and territorial performance in signaling male quality.

Characterizing Honeybee Cuticular Hydrocarbons During Foraging

Honeybees (Apis mellifera) adjust their time and effort during foraging activity. Their metabolic rates together with body temperature rise while gathering profitable resources. These physiological changes may result in a differential cuticular profile, which in turn may bear communicational value. We evaluated if sucrose concentration of collected food affects the cuticular chemistry of honeybees during foraging. We trained bees to artificial feeders with high (2 M) and low (0.5 M) sucrose concentrations, and captured the active foragers for surface extraction of cuticular compounds. We sampled foragers just after feeding, before taking-off towards the hive, and upon landing at the hive entrance, before entering the hive. Through gas chromatography-mass spectrometry analysis of cuticular extracts, we identified and quantified 48 compounds, including cuticular hydrocarbons (CHCs) and volatiles associated with exocrine glands. We found that higher sucrose concentrations resulted in increased amounts of alkanes and alkenes in the surface extracts of foragers captured at the hive entrance, but not at the feeding site. Our results suggest that the differences that have been reported for CHCs in waggle-dancing honey bees can be already found once they return to the hive from profitable food sources.