Behavioural consequences of intraspecific variability in a mate recognition signal.

Mate recognition is paramount for sexually reproducing animals, and many insects rely on cuticular hydrocarbons (CHCs) for close-range sexual communication. To ensure reliable mate recognition, intraspecific sex pheromone variability should be low. However, CHCs can be influenced by several factors, with the resulting variability potentially impacting sexual communication. While intraspecific CHC variability is a common phenomenon, the consequences thereof for mate recognition remain largely unknown. We investigated the effect of CHC variability on male responses in a parasitoid wasp showing a clear-cut within-population CHC polymorphism (three distinct female chemotypes, one thereof similar to male profiles). Males clearly discriminated between female and male CHCs, but not between female chemotypes in no-choice assays. When given a choice, a preference hierarchy emerged. Interestingly, the most attractive chemotype was the one most similar to male profiles. Mixtures of female CHCs were as attractive as chemotype-pure ones, while a female-male mixture negatively impacted male responses, indicating assessment of the entire, complex CHC profile composition. Our study reveals that the evaluation of CHC profiles can be strict towards 'undesirable' features, but simultaneously tolerant enough to cover a range of variants. This reconciles reliable mate recognition with naturally occurring variability.

Queens control caste allocation in the ant Cardiocondyla obscurior.

Social insect queens and workers can engage in conflict over reproductive allocation when they have different fitness optima. Here, we show that queens have control over queen-worker caste allocation in the ant Cardiocondyla obscurior, a species in which workers lack reproductive organs. We describe crystalline deposits that distinguish castes from the egg stage onwards, providing the first report of a discrete trait that can be used to identify ant caste throughout pre-imaginal development. The comparison of queen and worker-destined eggs and larvae revealed size and weight differences in late development, but no discernible differences in traits that may be used in social interactions, including hair morphology and cuticular odours. In line with a lack of caste-specific traits, adult workers treated developing queens and workers indiscriminately. Together with previous studies demonstrating queen control over sex allocation, these results show that queens control reproductive allocation in C. obscurior and suggest that the fitness interests of colony members are aligned to optimize resource allocation in this ant.

Cuticular Hydrocarbon Polymorphism in a Parasitoid Wasp.

Cuticular hydrocarbons (CHCs) are major constituents of the cuticular lipid layer of insects. They serve not only as a barrier to desiccation, but often additionally mediate communication at close range. The compositions of the CHC profiles, i.e., the specific compounds and their relative amounts, usually differ between species. Additional intraspecific variability can be found between different populations, between colonies and castes of social insects, and between the sexes. Thus, such groups can often be distinguished based on distinctive compounds and/or specific compound ratios. The CHC profile may further be influenced by biotic and abiotic factors, which therefore can impact, e.g., nestmate recognition or mate choice. However, consistent intrasexual variation seems to be rare. Here, we investigated a case of intrasexual CHC variability within a single population of a parasitoid wasp. While wasps of both sexes produced the same set of compounds, the relative amounts of specific compound classes revealed the presence of intrasexual chemical phenotypes. This is, to our knowledge, the first report of three distinct female CHC profile patterns within a population of a solitary insect that uses CHCs for mate recognition. Additionally, male CHC profiles, while overall very similar, could be separated into two chemotypes by multivariate analysis. The study of species exhibiting such intraspecific and intrasexual CHC variation will advance our understanding of the effects of CHC variability on both, desiccation resistance and intraspecific communication.

Social Behavior, Ovary Size, and Population of Origin Influence Cuticular Hydrocarbons in the Orchid Bee Euglossa dilemma.

AbstractCuticular hydrocarbons (CHCs) are waxy compounds on the surface of insects that prevent desiccation and frequently serve as chemical signals mediating social and mating behaviors. Although their function in eusocial species has been heavily investigated, little is known about the evolution of CHC-based communication in species with simpler forms of social organization lacking specialized castes. Here we investigate factors shaping CHC variation in the orchid bee Euglossa dilemma, which forms casteless social groups of two to three individuals. We first assess geographic variation, examining CHC profiles of males and females from three populations. We also consider CHC variation in the sister species, Euglossa viridissima, which occurs sympatrically with one population of E. dilemma. Next, we consider variation associated with female behavioral phases, to test the hypothesis that CHCs reflect ovary size and social dominance. We uncover a striking CHC polymorphism in E. dilemma spanning populations. In addition, we identify a separate set of CHCs that correlate with ovary size, social dominance, and expression of genes associated with social behavior, suggesting that CHCs convey reproductive and social information in E. dilemma. Together, our results reveal complex patterns of variation in which a subset of CHCs reflect the social and reproductive status of nestmates.

Parasitic wasps do not lack lipogenesis.

Fatty acids are crucial primary metabolites for virtually all creatures on earth. Most organisms thus do not rely exclusively on a nutritional supply containing fatty acids, but have the ability to synthesize fatty acids and triacylglycerides de novo from carbohydrates in a process called lipogenesis. The ubiquity of lipogenesis has been questioned by a series of studies reporting that many parasitic wasps (parasitoids) do not accumulate lipid mass despite having unlimited access to sugar. This has been interpreted as an evolutionary metabolic trait loss in parasitoids. Here, we demonstrate de novo biosynthesis of fatty acids from 13C-labelled α-d-glucose in 13 species of parasitoids from seven families. We furthermore show in the model organism Nasonia vitripennis that lipogenesis occurs even when lipid reserves are still intact, but relative 13C-incorporation rates increase in females with widely depleted fat reserves. We therefore conclude that the presumed 'lack of lipogenesis' in parasitoids needs to be re-evaluated.

De novo biosynthesis of linoleic acid is widespread in parasitic wasps.

Linoleic acid (C18:2∆9,12 , LA) is an important metabolite with numerous essential functions for growth, health, and reproduction of organisms. It has long been assumed that animals lack ∆12-desaturases, the enzymes needed to produce LA from oleic acid (C18:1∆9 , OA). There is, however, increasing evidence that this is not generally true for invertebrates. In the insect order Hymenoptera, LA biosynthesis has been shown for only two parasitic wasp species of the so-called "Nasonia group," but it is unknown whether members of other taxa are also capable of synthesizing LA. Here, we demonstrate LA biosynthesis in 13 out of 14 species from six families of parasitic wasps by gas chromatography-mass spectrometry analysis using two different stable isotope labeling techniques. Females of the studied species converted topically applied fully 13 C-labeled OA into LA and/or produced labeled LA after feeding on fully 13 C-labeled α- d-glucose. These results indicate that ∆12-desaturases are widespread in parasitic Hymenoptera and confirm previous studies demonstrating that these insects are capable of synthesizing fatty acids de novo.

Age-dependent release of and response to alarm pheromone in a ponerine ant.

Social insect societies are characterized by division of labour and communication within the colony. The most frequent mode of communication is by chemical signals. In general, pheromones elicit specific responses in the receiver, although reactions may vary depending on the receiving individual's physiological or motivational state. For example, it has been shown that pheromones can elicit different responses in morphological worker castes. However, comparably little is known about such effects in worker castes of monomorphic species. Here, we comprehensively studied a monomorphic species showing age polyethism, the thelytokous ant Platythyrea punctata Our analyses revealed that the species' alarm pheromone consists of (S)-(-)-citronellal and (S)-(-)-actinidine, and is produced in the mandibular glands. Ants responded with increased movement activity and increasing ant density towards the pheromone source in whole-colony bioassays, confirming the alarming effect of these compounds. We found age classes to differ in their absolute pheromone content, in the propensity to release alarm pheromone upon disturbance and in their reaction towards the pheromone. Absolute amounts of pheromone content may differ simply because the biosynthesis of the pheromone begins only after adult eclosion. Nonetheless, our results indicate that this clonal species exhibits age-related polyethism in the emission of as well as in the response to its alarm pheromone.

Blown by the wind: the ecology of male courtship display behavior in orchid bees.

Many insects rely on chemical signals to transmit precise information on the location, identity, and quality of potential mates. Chemical signals are often broadcasted at sites with physical properties that maximize signal propagation and signal transmission. Male neotropical orchid bees (Euglossini) perch and display on vertical branches and tree trunks in the forest to expose volatile blends (perfumes) that they previously collected from their environment. Previous studies have shown that the chemical composition of perfume blends is highly differentiated even between closely related species. However, variation in behavioral components of perfume exposure and male display remain poorly understood. We conducted a four-year study on orchid bee display sites (8 species) in pacific Costa Rica, using field observations along with chemical analysis and cage experiments to assess display niche partitioning among sympatric species. We evaluated the influence of physical factors (terrain, wind, light) on the distribution of perch sites and on display behavior, and tested a prediction of the sex pheromone-analogue hypothesis, i.e., that displaying males have above-average quantities or qualities of acquired perfumes. Males of different species displayed in the same general area and sometimes in close proximity to each other, but partitioned the display niche by selecting different perch diameters, display heights, and by displaying at different times of the day. Most perch sites were located inside the forest on elevated ground, especially along ridges, where stronger winds may help disperse perfume signals. Furthermore, the angular position of displaying males on perches was narrowly determined by wind direction, with males being positioned on the downwind side of the perch, where they would be most conspicuous to conspecifics approaching on an odor trail. Although our results generally support the hypothesis that perfumes serve as pheromone analogues, we did not find differences in the perfume composition of males caught at display perches and males captured at chemical baits. This suggests that, while chemical communication is an integral part of orchid bee display, male display activity is not determined by the history of, and success in, volatile acquisition.

Cuticular Hydrocarbons as Potential Close Range Recognition Cues in Orchid Bees.

Male Neotropical orchid bees collect volatile chemicals from their environment and compose species-specific volatile signals, which are subsequently exposed during courtship display. These perfumes are hypothesized to serve as attractants and may play a role in female mate choice. Here, we investigated the potential of cuticular hydrocarbons as additional recognition cues. The cuticular hydrocarbons of males of 35 species belonging to four of the five extant euglossine bee genera consisted of aliphatic hydrocarbons ranging in chain lengths between 21 and 37 C-atoms in distinct compositions, especially between sympatric species of similar coloring and size, for all but one case. Cleptoparasitic Exaerete spp. had divergent profiles, with major compounds predominantly constituted by longer hydrocarbon chains (>30 C-atoms), which may represent an adaptation to the parasitic life history ("chemical insignificance"). Phylogenetic comparative analyses imply that the chemical profiles exhibited by Exaerete spp. are evolutionarily divergent from the rest of the group. Female hydrocarbon profiles were not identical to male profiles in the investigated species, with either partial or complete separation between sexes in multivariate analyses. Sexually dimorphic hydrocarbon profiles are assumed to be the basis for sex recognition in a number of insects, and thus may supplement the acquired perfume phenotypes in chemical information transfer. Overall, cuticular hydrocarbons meet the requirements to function as intraspecific and intersexual close range recognition signals; behavioral experiments are needed to determine their potential involvement in mate recognition.

The Post-mating Switch in the Pheromone Response of Nasonia Females Is Mediated by Dopamine and Can Be Reversed by Appetitive Learning.

The olfactory sense is of crucial importance for animals, but their response to chemical stimuli is plastic and depends on their physiological state and prior experience. In many insect species, mating status influences the response to sex pheromones, but the underlying neuromodulatory mechanisms are poorly understood. After mating, females of the parasitic wasp Nasonia vitripennis are no longer attracted to the male sex pheromone. Here we show that this post-mating behavioral switch is mediated by dopamine (DA). Females fed a DA-receptor antagonist prior to mating maintained their attraction to the male pheromone after mating while virgin females injected with DA became unresponsive. However, the switch is reversible as mated females regained their pheromone preference after appetitive learning. Feeding mated N. vitripennis females with antagonists of either octopamine- (OA) or DA-receptors prevented relearning of the pheromone preference suggesting that both receptors are involved in appetitive learning. Moreover, DA injection into mated females was sufficient to mimic the oviposition reward during odor conditioning with the male pheromone. Our data indicate that DA plays a key role in the plastic pheromone response of N. vitripennis females and reveal some striking parallels between insects and mammals in the neuromodulatory mechanisms underlying olfactory plasticity.

Sublethal doses of imidacloprid disrupt sexual communication and host finding in a parasitoid wasp.

Neonicotinoids are widely used insecticides, but their use is subject of debate because of their detrimental effects on pollinators. Little is known about the effect of neonicotinoids on other beneficial insects such as parasitoid wasps, which serve as natural enemies and are crucial for ecosystem functioning. Here we show that sublethal doses of the neonicotinoid imidacloprid impair sexual communication and host finding in the parasitoid wasp Nasonia vitripennis. Depending on the dose, treated females were less responsive to the male sex pheromone or unable to use it as a cue at all. Courtship behaviour of treated couples was also impeded resulting in a reduction of mating rates by up to 80%. Moreover, treated females were no longer able to locate hosts by using olfactory cues. Olfaction is crucial for the reproductive success of parasitoid wasps. Hence, sublethal doses of neonicotinoids might compromise the function of parasitoid wasps as natural enemies with potentially dire consequences for ecosystem services.

Raising the sugar content--orchid bees overcome the constraints of suction feeding through manipulation of nectar and pollen provisions.

Unlike most other bees, the long-tongued orchid bees ingest nectar using suction feeding. Although long tongues allow exploitation of flowers with deep spurs, the energy intake rate is optimal at 10-20% lower nectar sugar concentrations compared to that of lapping bees. This constraint might be compensated by a higher digestive throughput. Additionally, orchid bees might evaporate water from regurgitated droplets of crop contents. We found male Euglossa championi (n = 10) and Euglossa dodsoni (n = 12) to regularly regurgitate droplets of crop content to the base of their proboscis, generating a fluid film between the proximal parts of the galeae, glossa and labial palps. Rhythmic movements of the proboscis may help to increase convection. There was a significant change in sugar concentration between the initially imbibed solution and the resulting crop content (P<0.05) and the time individual bees had engaged in this liquid exposure behavior was positively correlated with the resulting crop sugar concentration. Female Euglossa townsendi and Euglossa viridissima showed the same behavior. Additionally, they manipulated their nectar-enriched pollen provisions for extensive periods of time before deposition in brood cells. The deposited pollen loads (n = 14) showed a significantly higher sugar concentration than the sugar-water available to the bees (P<0.001). Thus, both male and female euglossines show behaviors that promote evaporative water loss from nectar. We suggest that the behaviors have evolved in concert with suction feeding on dilute nectar from deep floral tubes.