Cuticular Hydrocarbon Profiles Research Articles

Chemical Camouflage Induced by Diet in a Pest Treehopper on Host Plants.

Ants patrol foliage and exert a strong selective pressure on herbivorous insects, being their primary predators. As ants are chemically oriented, some organisms that interact with them (myrmecophiles) use chemical strategies mediated by their cuticular hydrocarbons (CHCs) to deal with ants. Thus, a better understanding of the ecology and evolution of the mutualistic interactions between myrmecophiles and ants depends on the accurate recognition of these chemical strategies. Few studies have examined whether treehoppers may use an additional strategy called chemical camouflage to reduce ant aggression, and none considered highly polyphagous pest insects. We analyzed whether the chemical similarity of the CHC profiles of three host plants from three plant families (Fabaceae, Malvaceae, and Moraceae) and the facultative myrmecophilous honeydew-producing treehopper Aetalion reticulatum (Hemiptera: Aetalionidae), a pest of citrus plants, may play a role as a proximate mechanism serving as a protection against ant attacks on plants. We found a high similarity (>80%) between the CHCs of the treehoppers and two of their host plants. The treehoppers acquire CHCs through their diet, and the chemical similarity varies according to host plant. Chemical camouflage on host plants plays a role in the interaction of treehoppers with their ant mutualistic partners.

Impact of diapause on the cuticular hydrocarbons and physiological status of Hippodamia variegata.

The spotted amber ladybird, Hippodamia variegata (Goeze) (Coleoptera: Coccinellidae), is known to be a potent predator of aphids, psyllids, whiteflies, mealybugs, and some butterfly species. This ladybeetle overwinters in the diapausing adult stage. The current study aimed to evaluate the impact of diapause on the energy resources and cuticular hydrocarbons (CHCs) of the female ladybeetle, specifically comparing the changes in glycogen, lipid, and protein contents, and CHCs profile of diapausing and non-diapausing adults. In this study, gas chromatography-mass was used to analyze whole-body extracts of the beetles. Results showed no significant differences between the amount of glycogen, lipid, and protein contents of diapausing and non-diapausing ladybeetle. The CHCs profile of H. variegata consisted of 24 hydrocarbons categorized into 2 groups: linear aliphatic hydrocarbons (n-alkanes) and methyl-branched hydrocarbons (17 molecules), as well as unsaturated cyclic compounds (7 molecules). The n-alkanes, with 14 compounds, were identified as the primary constituents of the CHCs of the ladybeetle. Six molecules were common to non-diapausing and diapausing beetles, 5 were exclusive to non-diapausing beetles, and 13 were exclusive to diapausing beetles. Moreover, we noted a significant difference in the quantity and quality of CHCs between diapausing and non-diapausing beetles, with diapausing beetles synthesizing more CHCs with longer chains. This disparity in CHC profiles was concluded to be an adaptation of H. variegata to survive harsh environmental conditions during diapause.

Targeted treatment of injured nestmates with antimicrobial compounds in an ant society

Infected wounds pose a major mortality risk in animals. Injuries are common in the ant Megaponera analis, which raids pugnacious prey. Here we show that M. analis can determine when wounds are infected and treat them accordingly. By applying a variety of antimicrobial compounds and proteins secreted from the metapleural gland to infected wounds, workers reduce the mortality of infected individuals by 90%. Chemical analyses showed that wound infection is associated with specific changes in the cuticular hydrocarbon profile, thereby likely allowing nestmates to diagnose the infection state of injured individuals and apply the appropriate antimicrobial treatment. This study demonstrates that M. analis ant societies use antimicrobial compounds produced in the metapleural glands to treat infected wounds and reduce nestmate mortality.

An experimental test of the evolutionary consequences of sympatry in Drosophila subquinaria.

Prezygotic isolation is often stronger between sympatric as opposed to allopatric taxa, but the underlying cause can be difficult to infer from comparative studies alone. Experimental evolution, where evolutionary responses to treatments manipulating the presence/absence of heterospecific individuals are tracked, can provide a powerful complementary approach. We used experimental evolution to investigate a naturally occurring pattern of reproductive character displacement in the mushroom-feeding fly, Drosophila subquinaria. In nature, female D. subquinaria from populations sympatric with the closely related Drosophila recens discriminate more strongly against heterospecific males than do females from allopatric populations. Starting with 16 replicate allopatric populations of D. subquinaria, we manipulated the presence/absence of D. recens during mating (experimental sympatry vs. control) and, when present, we allowed hybrids to live or kill them each generation. Across 12 generations, heterospecific offspring production from no-choice mating trials between D. subquinaria females and D. recens males declined in both experimental sympatry treatments relative to the control, suggesting increased sexual isolation. Male cuticular hydrocarbon profiles also evolved, but only in the hybrids killed treatment. Our results strongly imply that the existing reproductive character displacement in wild D. subquinaria populations was an evolutionary response to selection arising from secondary contact with D. recens.

Warming up through buildings and roads: what we know and should know about the urban heat island effect on bees

Urbanization leads to cities having higher temperatures than surrounding non-urban areas [this is known as the urban heat island (UHI) effect]. Very little is known about the impacts of the UHI effect on bees, despite the importance of temperature on many aspects of bees’ life suggesting that these may be not negligible. In this study, we aimed to highlight how the UHI effect could impact relevant functional traits of bees in cities, proposing several ad hoc hypotheses for traits that have thus far been investigated only in few studies or not at all, based on what we know from non-urban studies. The UHI effect was shown to influence bee body size, and generally tended to reduce the body size of bees in cities. Urban temperature may also affect bees’ wing morphology, and thus their overall flight morphology parameters. Individuals may be more brightly colored in cities. Bee ommatidial size and the number of antennal thermoreceptors they have may be smaller and fewer, respectively, in cities than in non-urban areas. As expected, because urban bees face a higher risk of desiccation, higher proportions of alkanes and longer main-carbon chain lengths are expected in their cuticular hydrocarbon (CHC) profiles. Stress biomarkers can also occur at greater concentrations in bees in cities and specific bacteria in the bee gut may occur at lower abundances. Warm urban temperatures may impact the life cycle of pathogens by reducing their proliferation. Aggression levels may be increased, and eusocial species may present more worker phases per year due to the UHI effect. All of these proposed impacts could be likely more visible in solitary and primitively eusocial bee species, which are those suspected to have a more limited dispersal ability. Comparative studies would help in the proper testing of these hypotheses.

Cuticular hydrocarbon profiles of Aedes aegypti reproductive stages - a potential tool for identifying high epidemiological risk populations

Cuticular hydrocarbon profiles of Aedes aegypti reproductive stages - a potential tool for identifying high epidemiological risk populations

Effects of genotype and host environment on the cuticular hydrocarbon profiles of Lysiphlebus parasitoids and aggression by aphid-tending ants.

Parasitoids in the genus Lysiphlebus specialize on ant-tended aphids and have previously been reported to mimic the cuticular hydrocarbon (CHC) profiles of their aphid hosts to avoid detection by ants. However, the precise mechanisms that mediate reduced ant aggression toward Lysiphlebus spp. are not known, nor is it clear whether such mechanisms are broadly effective or specialized on particular aphid hosts. Here we explore the effects of wasp genotype and host environment on Lysiphlebus CHC profiles and ant aggression. Rearing asexual Lysiphlebus lines in different host aphid environments revealed effects of both wasp line and aphid host on wasp CHCs. However, variation in genotype and host affected different features of the CHC profile, with wasp genotype explaining most variation in linear and long-chain methyl alkanes, while aphid host environment primarily influenced short-chain methyl alkanes. Subsequent behavioural experiments revealed no effects of host environment on ant aggression, but strong evidence for genotypic effects. The influence of genotypic variation on experienced ant aggression and relevant chemical traits is particularly relevant in light of recent evidence for genetic divergence among Lysiphlebus parasitoids collected from different aphid hosts.

Phylogenetic analyses of Reticulitermes (Blattodea: Rhinotermitidae) from California and other western states: multiple genes confirm undescribed species identified by cuticular hydrocarbons.

Subterranean termites in the genus Reticulitermes Holmgren 1913 are among the most economically important wood-destroying pests in the western United States. Yet, there remains uncertainty regarding the taxonomy and biology of the species in this genus. The 2 species described as having distributions in this region are the western subterranean termite, Reticulitermes hesperus Banks, and the arid land subterranean termite, Reticulitermes tibialis Banks. Taxonomic studies utilizing cuticular hydrocarbon (CHC) profiles, agonistic behavior, flight phenology, and mitochondrial DNA (mtDNA) suggested that R. hesperus is a species complex comprised of 2 or more sympatric, yet reproductively isolated species. To further delineate these taxa, we examined multiple genes from samples of Reticulitermes collected in the western United States. Alates collected after recent spring and fall mating flights, as well as previously collected workers, were subjected to CHC phenotyping and DNA sequence analyses that targeted mitochondrial cytochrome oxidase subunit II (COII), mitochondrial 16S rRNA, and nuclear Internal Transcribed Spacer 1 and 2 (ITS1 and 2). Phylogenetic analyses conducted also included published sequences of other putative western Reticulitermes species. Results suggest that at least 5 species of Reticulitermes may be present in California and that Reticulitermes in Arizona consistently group into multiple clades, including samples previously identified as R. tibialis in a sister clade. These analyses further support the species status of qualitatively different CHC phenotypes and that alates swarming in spring vs. fall are reproductively isolated species.

Chemical and population genetic analysis show no evidence of ecotype formation in a European population of the parasitoid wasp Nasonia vitripennis

Ecotypes, subpopulations or strains of a single species locally adapted to divergent ecological conditions within the same habitat are often considered to be the first steps in sympatric speciation. It has been suggested that two ecotypes are distinguishable in Nasonia vitripennis, a prominent model organism for parasitic Hymenoptera, with one ecotype parasitizing fly pupae in bird nests, and the other one parasitizing fly pupae on carrion. This differentiation into two ecotypes has been hypothesized to indicate incipient sympatric speciation in populations of this globally distributed species. In the present study, we investigated the differentiation into these two distinct ecotypes focusing on chemical profiles and the population genetic divergence in a wild N. vitripennis population from the Netherlands. Isofemale lines were obtained from bird nest boxes and from deer carrion, respectively, representing both microhabitats. To test for phenotypic differentiation, we determined the surface cuticular hydrocarbon (CHC) profiles from wasps of both host patches. Using a panel of 14 microsatellites, we concordantly determined the population genetic structure and tested for genetic differentiation between foundresses obtained from both microhabitats. Both the phenotypic as well as the genetic datasets show no evidence for any kind of separation based on the postulated two ecotypes, but rather suggest free interbreeding with no gene flow interruption between the two distinct host patches. Our findings challenge previous assumptions on clearly distinguishable ecotypes in N. vitripennis, and demonstrate how a chemical ecological assessment coupled with population genetics can be instrumental in re-evaluating the potential of ecological differentiation and incipient speciation mechanisms in parasitoid wasps.

A predatory social wasp does not avoid nestmates contaminated with a fungal biopesticide.

Fungus-based biopesticides have been used worldwide for crop pest control as a safer alternative to chemical pesticides such as neonicotinoids. Both agrochemicals can be lethal and may also trigger side effects on the behavioral traits of non-target social insects, which play a crucial role in providing essential biological pest control services in agroecosystems. Here, we evaluated whether a commercial formulation of the entomopathogenic fungus Beauveria bassiana or the neonicotinoid imidacloprid causes mortality in foragers of Mischocyttarus metathoracicus. These social wasps are natural enemies of caterpillars and other herbivorous insects and inhabit both urban and agricultural environments in Brazil. We also tested whether wasps discriminate between biopesticide-exposed and unexposed conspecifics. Through a combination of laboratory (survival assay) and field experiments (lure presentation), along with chemical analyses (cuticular hydrocarbon profiles), we showed that topic exposure to the label rate of each pesticide causes a lethal effect, with the biopesticide exhibiting a slower effect. Moreover, wasps do not discriminate biopesticide-exposed from unexposed conspecifics, likely because of the similarity of their cuticular chemical profiles 24h after exposure. Overall, the delayed lethal time at the individual level, combined with the indistinctive chemical cues of exposure and the lack of discrimination by conspecifics suggests that the fungal biopesticide may ultimately pose a threat to the colony survival of this predatory wasp.

Exposure to sublethal concentration of flupyradifurone alters sexual behavior and cuticular hydrocarbon profile in Heriades truncorum, an oligolectic solitary bee.

The aboveground oligolectic bee, Heriades truncorum, is a particularly good model for studying the impact of pesticides on sexual communication, since some aspects of its mating behavior have previously been described. We have tested (1) the interference of the pesticide flupyradifurone on male precopulatory behavior and male mating partner preferences, (2) the way that the pesticide interferes in male quality assessment by the female, and (3) the effects of the pesticide on the chemical compounds in the female cuticle. We exposed bees of both sexes to a sublethal concentration of flupyradifurone. Various behaviors were registered in a mating arena with two females (one unexposed and one exposed) and one male (either unexposed or exposed). Unexposed males were quicker to attempt to mate. Treatment also impacted precopulatory behavior and male quality assessment by females. Males approached unexposed females more quickly than insecticide-exposed ones. Females exposed to insecticide produced lower amounts of some cuticular hydrocarbons (sex pheromone candidates) and appeared less choosy than unexposed females. Our findings suggest that insecticide exposure affects sexual communication, playing a role both in male preference and in male quality assessment by the female.

Cuticular Hydrocarbon Profiling of Australian Gonipterini Weevils

Cuticular hydrocarbon (CHC) profiling shows promise as a chemotaxonomic tool for identifying and discriminating between closely related insect species. However, there have been limited studies using CHC profiling to differentiate between weevil species (Coleoptera: Curculionidae). This proof-of-concept study investigated the use of CHC and volatile profiling to discriminate between five weevil species from three genera in the Gonipterini tribe. A total of 56 CHCs and 41 other volatile compounds were found across the five species, with 83 of the compounds being identified through their mass fragmentation patterns. The number of CHCs from each species ranged from 20 to 43, while the proportion of CHCs unique to each species varied between 0% and 19%. The most abundant CHCs were nonacosane, 7-methylheptacosane, heptacosane, and hexacosane. Principal component analysis of the centred log-ratio transformed data revealed broad differences in CHC profiles between the two Oxyops species, with Bryachus squamicollis demonstrating the greatest divergence from the other Gonipterini species. The results suggest that CHC analysis could be used to support established taxonomic methods, including morphological features and genetic sequencing results.

Male exposure to guava volatiles affects female remating behavior in Anastrepha fraterculus

AbstractPlant‐derived volatiles can have positive or negative effects on insects. Exposure to volatiles of guava (Psidium guajava L.) (Myrtaceae) enhances male mating success in the South American fruit fly, Anastrepha fraterculus (Wiedemann) (Diptera: Tephritidae), an important pest of various fruits. These volatiles can therefore be used as a post‐teneral, pre‐release supplement in sterile insect technique (SIT) programs. In SIT, mass‐reared sterile males compete for and mate with wild females, in order to reduce populations of insect pests. However, the success of SIT will be diminished if males are not able to inhibit females from remating with other (potentially wild) males. Here, we investigated the remating propensity of A. fraterculus females mated with males exposed to guava volatiles or non‐exposed (control) in choice and no‐choice tests. In addition, we tested whether exposure to guava volatiles affected male reproductive organ size (testes, accessory glands, and seminal vesicles) and cuticular hydrocarbon (CHC) profile. Females mated with guava‐exposed males were more likely to remate than females mated with control males, but there was no difference in the length of the refractory period (i.e., the time between the first and second mating event). Reproductive organ size did not differ between exposed and non‐exposed males, but exposed males had a higher quantity of waxes than non‐exposed males, and CHC profiles differed. The results are discussed in the context of SIT programs for the control of this insect pest.

Queen fecundity, worker entourage and cuticular chemistry in the ant Formica fusca.

Cooperative breeding entails conflicts over reproductive shares that may be settled in different ways. In ants, where several queens simultaneously reproduce in a colony, both queens and workers may influence the reproductive apportionment and offspring quality. Queens may vary in their intrinsic fecundity, which may influence the size of the worker entourage attending individual queens, and this may eventually dictate the reproductive output of a queen. We tested whether the reproductive success of queens is affected by the size of their worker entourage, their fecundity at the onset of the reproductive season, and whether the queen cuticular hydrocarbon profile carries information on fecundity. We show that in the ant Formica fusca both queen fecundity and egg hatching success increase with the size of their entourage, and that newly hatched larvae produced by initially highly fecund queens are smaller. Furthermore, higher relatedness among workers increased queen fecundity. Finally, the queens that received a large worker entourage differed in the cuticular chemistry from those that received a small worker entourage. Our results thus show that workers play a pivotal role in determining queen fitness, that high intracolony relatedness among workers enhances the overall reproductive output in the colony, and that queen fecundity is reflected in their cuticular hydrocarbon profile.

Three cuticular amides in the tripartite symbiosis of leafcutter ants.

Cuticular hydrocarbons (CHCs) play various roles in insects' chemical ecology. As leafcutter ants live in a specific symbiosis with fungi, they harvest and with different bacteria, some of these CHCs might be associated with a mutualistic function within this symbiosis. To obtain a more precise picture in that respect we compared the CHC profiles of the leafcutter ants, Atta sexdens, Atta cephalotes, and Acromyrmex octospinosus inhabited by mutualistic bacteria with the profiles of Polyrhachis dives and Messor aciculatus by GC-EI-MS analysis and 28 other ant species by data from the literature. We were able to identify three alkyl amides (hexadecanamide, hexadecenamide, and tetradecanamide), occurring only in the CHC profiles of leafcutter ants inhabited by symbiotic bacteria. Our results lead to the conclusion that those alkyl amides could have a function in the tripartite symbiosis of leafcutter ants.

Humidity stress and its consequences for male pre‐ and post‐copulatory fitness traits in an insect

AbstractGlobal declines in insect abundance are of significant concern. While there is evidence that climate change is contributing to insect declines, we know little of the direct mechanisms responsible for these declines. Male fertility is compromised by increasing temperatures, and the thermal limit to fertility has been implicated as an important factor in the response of insects to climate change. However, climate change is affecting both temperature and hydric conditions, and the effects of water availability on male fertility have rarely been considered. Here we exposed male crickets Teleogryllus oceanicus to either low or high‐humidity environments while holding temperature constant. We measured water loss and the expression of both pre‐ and postmating reproductive traits. Males exposed to a low‐humidity environment lost more water than males exposed to a high‐humidity environment. A male's cuticular hydrocarbon profile (CHC) did not affect the amount of water lost, and males did not adjust the composition of their CHC profiles in response to hydric conditions. Males exposed to a low‐humidity environment were less likely to produce courtship song or produced songs of low quality. Their spermatophores failed to evacuate and their ejaculates contained sperm of reduced viability. The detrimental effects of low‐humidity on male reproductive traits will compromise male fertility and population persistence. We argue that limits to insect fertility based on temperature alone are likely to underestimate the true effects of climate change on insect persistence and that the explicit incorporation of water regulation into our modeling will yield more accurate predictions of the effects of climate change on insect declines.

Integrating lipid metabolism, pheromone production and perception by Fruitless and Hepatocyte Nuclear Factor 4.

Sexual attraction and perception are crucial for mating and reproductive success. In Drosophila melanogaster, the male-specific isoform of Fruitless (Fru), FruM, is a known master neuro-regulator of innate courtship behavior to control the perception of sex pheromones in sensory neurons. Here, we show that the non-sex-specific Fru isoform (FruCOM) is necessary for pheromone biosynthesis in hepatocyte-like oenocytes for sexual attraction. Loss of FruCOM in oenocytes resulted in adults with reduced levels of cuticular hydrocarbons (CHCs), including sex pheromones, and show altered sexual attraction and reduced cuticular hydrophobicity. We further identify Hepatocyte nuclear factor 4 (Hnf4) as a key target of FruCOM in directing fatty acid conversion to hydrocarbons. Fru or Hnf4 depletion in oenocytes disrupts lipid homeostasis, resulting in a sex-dimorphic CHC profile that differs from doublesex- and transformer-dependent CHC dimorphism. Thus, Fru couples pheromone perception and production in separate organs to regulate chemosensory communications and ensure efficient mating behavior.

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.

Conserved worker policing in African carpenter ants with drastically different egg chemotypes

Distinctive chemical signatures have the potential to serve as discriminatory cues for olfactory recognition mechanisms. Cuticular hydrocarbon (CHC) profiles are among the most prominent chemical signatures in insects that can be highly diverse even among closely related species and between populations with similar ecology. Particularly within the major insect order Hymenoptera, CHC profiles are characterized by high complexity and variation with the potential to evolve rapidly. In this study, we found two very distinct CHC chemotypes distinguishing sympatric colonies of the African carpenter ant Camponotus maculatus (Hymenoptera: Formicinae). These chemotypic differences were mainly detected on the surface profiles of eggs produced by either queens or isolated worker groups. In one chemotype, queen- and worker-laid eggs are very similar. This is largely contrasted by the other chemotype, where queen-laid eggs clearly differ from worker-laid eggs with several prominent queen-exclusive compounds. However, workers display a stable behavior of discriminating against and selectively disposing of worker-laid eggs i.e., worker policing, independent of egg chemotype. Furthermore, genetic barcoding of workers revealed a clear separation between colonies characterized by producing these two distinct egg chemotypes, which may indicate that these colonies belong to a cryptic species complex. Interestingly, worker policing behaviour appears to be evolutionarily conserved, despite the strikingly different egg surface profiles.

Evidence of cuticular hydrocarbon profile alterations by Wolbachia in females, but not males, of an aculeate parasitoid wasp

Evidence of cuticular hydrocarbon profile alterations by Wolbachia in females, but not males, of an aculeate parasitoid wasp