Floral scent of the Mediterranean fig tree: significant inter-varietal difference but strong conservation of the signal responsible for pollinator attraction.

For thousands of years, humans have domesticated different plants by selecting for particular characters, often affecting less-known traits, including the volatile organic compounds (VOCs) emitted by these plants for defense or reproduction. The fig tree Ficus carica has a very wide range of varieties in the Mediterranean region and is selected for its traits affecting fruits, including pollination, but the effect of human-driven diversification on the VOCs emitted by the receptive figs to attract their pollinator (Blastophaga psenes) is not known. In the present study, VOCs from receptive figs of eight varieties in northern Morocco, were collected at different times within the manual pollination period and analyzed by gas chromatography-mass spectrometry. Genetic analyses using microsatellite loci were performed on the same varieties. Despite strong inter-varietal differences in the quantity and relative proportions of all VOCs, the relative proportions of the four pollinator-attractive VOCs showed limited variation among varieties. There was no significant correlation between genetic markers and chemical profiles of the different varieties. While diversification driven by humans has led to differences between varieties in VOC profiles, this paper suggests that throughout the process of domestication and varietal diversification, stabilizing selection has maintained a strong signal favoring pollinator attraction.

Extensive human-mediated jump dispersal within and across the native and introduced ranges of the invasive termite Reticulitermes flavipes.

As native ranges are often geographically structured, invasive species originating from a single source population only carry a fraction of the genetic diversity present in their native range. The invasion process is thus often associated with a drastic loss of genetic diversity resulting from a founder event. However, the fraction of diversity brought to the invasive range may vary under different invasion histories, increasing with the size of the propagule, the number of reintroduction events, and/or the total genetic diversity represented by the various source populations in a multiple-introduction scenario. In this study, we generated a SNP data set for the invasive termite Reticulitermes flavipes from 23 native populations in the eastern United States and six introduced populations throughout the world. Using population genetic analyses and approximate Bayesian computation random forest, we investigated its worldwide invasion history. We found a complex invasion pathway with multiple events out of the native range and bridgehead introductions from the introduced population in France. Our data suggest that extensive long-distance jump dispersal appears common in both the native and introduced ranges of this species, probably through human transportation. Overall, our results show that similar to multiple introduction events into the invasive range, admixture in the native range prior to invasion can potentially favour invasion success by increasing the genetic diversity that is later transferred to the introduced range.

Strong Gene Flow Undermines Local Adaptations in a Host Parasite System.

The co-evolutionary pathways followed by hosts and parasites strongly depend on the adaptive potential of antagonists and its underlying genetic architecture. Geographically structured populations of interacting species often experience local differences in the strength of reciprocal selection pressures, which can result in a geographic mosaic of co-evolution. One example of such a system is the boreo-montane social wasp Polistes biglumis and its social parasite Polistes atrimandibularis, which have evolved local defense and counter-defense mechanisms to match their antagonist. In this work, we study spatial genetic structure of P. biglumis and P. atrimandibularis populations at local and regional scales in the Alps, by using nuclear markers (DNA microsatellites, AFLP) and mitochondrial sequences. Both the host and the parasite populations harbored similar amounts of genetic variation. Host populations were not genetically structured at the local scale, but geographic regions were significantly differentiated from each other in both the host and the parasite in all markers. The net dispersal inferred from genetic differentiation was similar in the host and the parasite, which may be due to the annual migration pattern of the parasites between alpine and lowland populations. Thus, the apparent dispersal barriers (i.e., high mountains) do not restrict gene flow as expected and there are no important gene flow differences between the species, which contradict the hypothesis that restricted gene flow is required for local adaptations to evolve.

Invasion Dynamics of A Termite, Reticulitermes flavipes, at Different Spatial Scales in France.

Termites are social insects that can also be major pests. A well-known problem species is the subterranean termite, Reticulitermes flavipes. It is invasive in France and is thought to have arrived from Louisiana during the 18th century. While the putative source of French populations has been identified, little is known about how the termite spread following its establishment. Here, we examined expansion patterns at different spatial scales in urban areas to clarify how R. flavipes spread in France. Based on our analyses of phylogeography and population genetics, results suggest a scenario of successive introductions into the Charente-Maritime region, on the Atlantic Coast. Two major expansion fronts formed: one that spread toward the northeast and the other toward the southeast. At the regional scale, different spatial and genetic distribution patterns were observed: there was heterogeneity in Île-de-France and aggregation in Centre-Val de Loire. At the local scale, we found that our three focal urban sites each formed a single large colony that contained several secondary reproductives. Our findings represent a second step in efforts to reconstruct termite's invasion dynamics. They also highlight the role that may have been played by the French railway network in transporting termites over long distances.

Unbalanced biparental care during colony foundation in two subterranean termites.

Parental care is a major component of reproduction in social organisms, particularly during the foundation steps. Because investment into parental care is often costly, each parent is predicted to maximize its fitness by providing less care than its partner. However, this sexual conflict is expected to be low in species with lifelong monogamy, because the fitness of each parent is typically tied to the other's input. Somewhat surprisingly, the outcomes of this tug-of-war between maternal and paternal investments have received important attention in vertebrate species, but remain less known in invertebrates. In this study, we investigated how queens and kings share their investment into parental care and other social interactions during colony foundation in two termites with lifelong monogamy: the invasive species Reticulitermes flavipes and the native species R. grassei. Behaviors of royal pairs were recorded during six months using a non-invasive approach. Our results showed that queens and kings exhibit unbalanced investment in terms of grooming, antennation, trophallaxis, and vibration behavior. Moreover, both parents show behavioral differences toward their partner or their descendants. Our results also revealed differences among species, with R. flavipes exhibiting shorter periods of grooming and antennation toward eggs or partners. They also did more stomodeal trophallaxis and less vibration behavior. Overall, this study emphasizes that despite lifelong monogamy, the two parents are not equally involved in the measured forms of parental care and suggests that kings might be specialized in other tasks. It also indicates that males could play a central, yet poorly studied role in the evolution and maintenance of the eusocial organization.

Nest signature changes throughout colony cycle and after social parasite invasion in social wasps.

Social insects recognize their nestmates by means of a cuticular hydrocarbon signature shared by colony members, but how nest signature changes across time has been rarely tested in longitudinal studies and in the field. In social wasps, the chemical signature is also deposited on the nest surface, where it is used by newly emerged wasps as a reference to learn their colony odor. Here, we investigate the temporal variations of the chemical signature that wasps have deposited on their nests. We followed the fate of the colonies of the social paper wasp Polistes biglumis in their natural environment from colony foundation to decline. Because some colonies were invaded by the social parasite Polistes atrimandibularis, we also tested the effects of social parasites on the nest signature. We observed that, as the season progresses, the nest signature changed; the overall abundance of hydrocarbons as well as the proportion of longer-chain and branched hydrocarbons increased. Where present, social parasites altered the host-nest signature qualitatively (adding parasite-specific alkenes) and quantitatively (by interfering with the increase in overall hydrocarbon abundance). Our results show that 1) colony odor is highly dynamic both in colonies controlled by legitimate foundresses and in those controlled by social parasites; 2) emerged offspring contribute little to colony signature, if at all, in comparison to foundresses; and 3) social parasites, that later mimic host signature, initially mark host nests with species-specific hydrocarbons. This study implies that important updating of the neural template used in nestmate recognition should occur in social insects.

An American termite in Paris: temporal colony dynamics.

Termites of the genus Reticulitermes are widespread invaders, particularly in urban habitats. Their cryptic and subterranean lifestyle makes them difficult to detect, and we know little about their colony dynamics over time. In this study we examined the persistence of Reticulitermes flavipes (Kollar) colonies in the city of Paris over a period of 15 years. The aim was (1) to define the boundaries of colonies sampled within the same four areas over two sampling periods, (2) to determine whether the colonies identified during the first sampling period persisted to the second sampling period, and (3) to compare the results obtained when colonies were delineated using a standard population genetic approach versus a Bayesian clustering method that combined both spatial and genetic information. Herein, colony delineations were inferred from genetic differences at nine microsatellite loci and one mitochondrial locus. Four of the 18 identified colonies did not show significant differences in their genotype distributions between the two sampling periods. While allelic richness was low, making it hard to reliably distinguish colony family type, most colonies appeared to retain the same breeding structure over time. These large and expansive colonies showed an important ability to fuse (39% were mixed-family colonies), contained hundreds of reproductives and displayed evidence of isolation-by-distance, suggesting budding dispersal. These traits, which favor colony persistence over time, present a challenge for pest control efforts, which apply treatment locally. The other colonies showed significant differences, but we cannot exclude the possibility that their genotype distributions simply changed over time.

Lock-picks: fungal infection facilitates the intrusion of strangers into ant colonies.

Studies investigating host-parasite systems rarely deal with multispecies interactions, and mostly explore impacts on hosts as individuals. Much less is known about the effects at colony level, when parasitism involves host organisms that form societies. We surveyed the effect of an ectoparasitic fungus, Rickia wasmannii, on kin-discrimination abilities of its host ant, Myrmica scabrinodis, identifying potential consequences at social level and subsequent changes in colony infiltration success of other organisms. Analyses of cuticular hydrocarbons (CHCs), known to be involved in insects' discrimination processes, revealed variations in chemical profiles correlated with the infection status of the ants, that could not be explained by genetic variation tested by microsatellites. In behavioural assays, fungus-infected workers were less aggressive towards both non-nestmates and unrelated queens, enhancing the probability of polygyny. Likewise, parasitic larvae of Maculinea butterflies had a higher chance of adoption by infected colonies. Our study indicates that pathogens can modify host recognition abilities, making the society more prone to accept both conspecific and allospecific organisms.

Subterranean termite phylogeography reveals multiple postglacial colonization events in southwestern Europe.

A long-standing goal of evolutionary biology is to understand how paleoclimatic and geological events shape the geographical distribution and genetic structure within and among species. Using a diverse set of markers (cuticular hydrocarbons, mitochondrial and nuclear gene sequences, microsatellite loci), we studied Reticulitermes grassei and R. banyulensis, two closely related termite species in southwestern Europe. We sought to clarify the current genetic structure of populations that formed following postglacial dispersal from refugia in southern Spain and characterize the gene flow between the two lineages over the last several million years. Each marker type separately provided a fragmented picture of the evolutionary history at different timescales. Chemical analyses of cuticular hydrocarbons and phylogenetic analyses of mitochondrial and nuclear genes showed clear separation between the species, suggesting they diverged following vicariance events in the Late Miocene. However, the presence of intermediate chemical profiles and mtDNA introgression in some Spanish colonies suggests ongoing gene flow. The current genetic structure of Iberian populations is consistent with alternating isolation and dispersal events during Quaternary glacial periods. Analyses of population genetic structure revealed postglacial colonization routes from southern Spain to France, where populations underwent strong genetic bottlenecks after traversing the Pyrenees resulting in parapatric speciation.

Historical biogeography of Reticulitermes termites (Isoptera: Rhinotermitidae) inferred from analyses of mitochondrial and nuclear loci.

Termites of the genus Reticulitermes are ecologically and economically important wood-feeding social insects that are widespread in the Holarctic region. Despite their importance, no study has yet attempted to reconstruct a global time-scaled phylogeny of Reticulitermes termites. In this study, we sequenced mitochondrial (2096bp) and nuclear (829bp) loci from 61 Reticulitermes specimens, collected across the genus' entire range, and one specimen of Coptotermes formosanus, which served as an outgroup. Bayesian and Maximum likelihood analyses conducted on the mitochondrial and nuclear sequences support the existence of four main lineages that span four global geographical regions: North America (NA lineage), western Europe (WE lineage), a region including eastern Europe and western Asia (EA+WA lineage), and eastern Asia (EA lineage). The mitochondrial data allowed us to clarify the phylogenetic relationships among these lineages. They were also used to infer a chronogram that was time scaled based on age estimates for termite fossils (including the oldest Reticulitermes fossils, which date back to the late Eocene-early Oligocene). Our results support the hypothesis that the extant Reticulitermes lineage first differentiated in North America. The first divergence event in the ancestral lineage of Reticulitermes occurred in the early Miocene and separated the Nearctic lineages (i.e., the NA lineages) from the Palearctic lineages (i.e., WE, EE+WA, and EA lineages). Our analyses revealed that the main lineages of Reticulitermes diversified because of vicariance and migration events, which were probably induced by major paleogeographic and paleoclimatic changes that occurred during the Cenozoic era. This is the first global and comprehensive phylogenetic study of Reticulitermes termites, and it provides a crucial foundation for studying the evolution of phenotypic and life-history traits in Reticulitermes. For instance, the phylogeny we obtained suggested that 'asexual queen succession', a unique reproductive system, independently evolved at least three times during the diversification of the genus.

Spatial structuring of the population genetics of a European subterranean termite species.

In population genetics studies, detecting and quantifying the distribution of genetic variation can help elucidate ecological and evolutionary processes. In social insects, the distribution of population-level genetic variability is generally linked to colony-level genetic structure. It is thus especially crucial to conduct complementary analyses on such organisms to examine how spatial and social constraints interact to shape patterns of intraspecific diversity. In this study, we sequenced the mitochondrial COII gene for 52 colonies of the subterranean termite Reticulitermes grassei (Isoptera: Rhinotermitidae), sampled from a population in southwestern France. Three haplotypes were detected, one of which was found exclusively in the southern part of the study area (near the Pyrenees). After genotyping 6 microsatellite loci for 512 individual termites, we detected a significant degree of isolation by distance among individuals over the entire range; however, the cline of genetic differentiation was not continuous, suggesting the existence of differentiated populations. A spatial principal component analysis based on allele frequency data revealed significant spatial autocorrelation among genotypes: the northern and southern groups were strongly differentiated. This finding was corroborated by clustering analyses; depending on the randomized data set, two or three clusters, exhibiting significant degrees of differentiation, were identified. An examination of colony breeding systems showed that colonies containing related neotenic reproductives were prevalent, suggesting that inbreeding may contribute to the high level of homozygosity observed and thus enhance genetic contrasts among colonies. We discuss the effect of evolutionary and environmental factors as well as reproductive and dispersal modes on population genetic structure.

Population diversity in cuticular hydrocarbons and mtDNA in a mountain social wasp.

Nestmate recognition is a common phenomenon in social insects that typically is mediated by cuticular hydrocarbons. Geographical variation in cuticular hydrocarbons has been observed, although the pattern of variation is not consistent across species and is usually related to the biology and ecology of the different species. Polistes biglumis (Hymenoptera: Vespidae) is a social wasp that lives in high mountains where populations are separated by significant geographical barriers. Here we investigated the level of chemical variation among populations of P. biglumis in the Alps, and shed light on the phylogeography of this species. Populations could be discriminated by means of their cuticular hydrocarbon profiles, which showed a pattern consistent with the isolation-by-distance hypothesis. Molecular data highlighted two areas with different levels of haplotype diversity, although all wasps belonged to the same species. These results suggest that the populations of P. biglumis in the Alps are geographically isolated from one another, favoring their genetic and chemical differentiation.

Colony breeding structure of the invasive termite Reticulitermes urbis (Isoptera: Rhinotermitidae).

Invasive species cause severe environmental and economic problems. The invasive success of social insects often appears to be related to their ability to adjust their social organization to new environments. To gain a better understanding of the biology of invasive termites, this study investigated the social organization of the subterranean termite, Reticulitermes urbis, analyzing the breeding structure and the number of reproductives within colonies from three introduced populations. By using eight microsatellite loci to determine the genetic structure, it was found that all the colonies from the three populations were headed by both primary reproductives (kings and queens) and secondary reproductives (neotenics) to form extended-family colonies. R. urbis appears to be the only Reticulitermes species with a social organization based solely on extended-families in both native and introduced populations, suggesting that there is no change in their social organization on introduction. F-statistics indicated that there were few neotenics within the colonies from urban areas, which did not agree with results from previous studies and field observations. This suggests that although several neotenics may be produced, only few become active reproductives. The results also imply that the invasive success of R. urbis may be based on different reproductive strategies in urban and semiurbanized areas. The factors influencing an individual to differentiate into a neotenic in Reticulitermes species are discussed.

Changes in the hydrocarbon proportions of colony odor and their consequences on nestmate recognition in social wasps.

In social insects, colonies have exclusive memberships and residents promptly detect and reject non-nestmates. Blends of epicuticular hydrocarbons communicate colony affiliation, but the question remains how social insects use the complex information in the blends to discriminate between nestmates and non-nestmates. To test this we altered colony odor by simulating interspecific nest usurpation. We split Polistes dominulus paper-wasp nests into two halves and assigned a half to the original foundress and the other half to a P. nimphus usurper for 4 days. We then removed foundresses and usurpers from nests and investigated whether emerging P. dominulus workers recognized their never-before-encountered mothers, usurpers and non-nestmates of the two species. Behavioral and chemical analyses of wasps and nests indicated that 1) foundresses marked their nests with their cuticular hydrocarbons; 2) usurpers overmarked foundress marks and 3) emerging workers learned colony odor from nests as the odor of the female that was last on nest. However, notwithstanding colony odor was usurper-biased in usurped nests, workers from these nests recognized their mothers, suggesting that there were pre-imaginal and/or genetically encoded components in colony-odor learning. Surprisingly, workers from usurped nests also erroneously tolerated P. nimphus non-nestmates, suggesting they could not tell odor differences between their P. nimphus usurpers and P. nimphus non-nestmates. Usurpers changed the odors of their nests quantitatively, because the two species had cuticular hydrocarbon profiles that differed only quantitatively. Possibly, P. dominulus workers were unable to detect differences between nestmate and non-nestmate P. nimphus because the concentration of some peaks in these wasps was beyond the range of workers' discriminatory abilities (as stated by Weber's law). Indeed, workers displayed the least discrimination abilities in the usurped nests where the relative odor changes due to usurpation were the largest, suggesting that hydrocarbon variations beyond species-specific ranges can alter discrimination abilities.

Clinal variation in colony breeding structure and level of inbreeding in the subterranean termites Reticulitermes flavipes and R. grassei.

Social insects exhibit remarkable variation in their colony breeding structures, both within and among species. Ecological factors are believed to be important in shaping reproductive traits of social insect colonies, yet there is little information linking specific environmental variables with differences in breeding structure. Subterranean termites (Rhinotermitidae) show exceptional variation in colony breeding structure, differing in the number of reproductives and degree of inbreeding; colonies can be simple families headed by a single pair of monogamous reproductives (king and queen) or they can be extended families headed by multiple inbreeding neotenic reproductives (wingless individuals). Using microsatellite markers, we characterized colony breeding structure and levels of inbreeding in populations over large parts of the range of the subterranean termites Reticulitermes flavipes in the USA and R. grassei in Europe. Combining these new data with previous results on populations of both species, we found that latitude had a strong effect on the proportion of extended-family colonies in R. flavipes and on levels of inbreeding in both species. We examined the effect of several environmental variables that vary latitudinally; while the degree of inbreeding was greatest in cool, moist habitats in both species, seasonality affected the species differently. Inbreeding in R. flavipes was most strongly associated with climatic variables (mean annual temperature and seasonality), whereas nonclimatic variables, including the availability of wood substrate and soil composition, were important predictors of inbreeding in R. grassei. These results are the first showing that termite breeding structure is shaped by local environmental factors and that species can vary in their responses to these factors.

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.

Brood care and social evolution in termites.

Cooperative brood care is assumed to be the common driving factor leading to sociality. While this seems to be true for social Hymenoptera and many cooperatively breeding vertebrates, the importance of brood care for the evolution of eusociality in termites is unclear. A first step in elucidating this problem is an assessment of the ancestral condition in termites. We investigated this by determining the overall level of brood care behaviour across four termite species that cover the phylogenetic diversity of the lower termites. Brood care was low in the three species (all from different families) that had an ancestral wood-dwelling lifestyle of living in a single piece of wood that serves as food and shelter. In the fourth species, a lower termite that evolved outside foraging, brood care was more common. Together with data for higher termites, this suggests that brood care in termites only becomes important when switching from a wood-dwelling to a foraging lifestyle. These results imply that early social evolution in termites was driven by benefits of increased defence, while eusociality in Hymenoptera and cooperative breeding in birds and mammals are primarily based on brood care.

Ontogenic potentialities of the worker caste in two sympatric subterranean termites in France.

In termites, the capacity of workers to differentiate into neotenic reproductives is an important characteristic that deserves particular attention. To gain insight into the differentiation pathway, the potentialities of workers and the endocrinal changes during the formation of neotenics were compared in two sympatric termites, Reticulitermes flavipes and Reticulitermes grassei. After 1 year of development, 100% of R. flavipes worker groups produced neotenics against only 63% of R. grassei groups. The average production of female neotenics was significantly higher in R. flavipes worker groups compared with R. grassei groups and R. flavipes produced a greater proportion of female neotenics. Moreover, R. flavipes produced more offspring, not only because there were more females, but also because R. flavipes females were more productive. Moreover, the offspring produced by R. flavipes grew faster than the offspring of R. grassei. Both ecdysteroid and juvenile hormone (JH) titers varied significantly during the development of neotenics. The two species showed similar ecdysteroid titer variation patterns. However, the JH titer variation patterns strongly differed: in R. grassei, the concentration of JH increased in maturing neotenics then dropped in mature neotenics, whereas in R. flavipes, the level of JH was significantly higher than in R. grassei and remained constantly high in mature neotenics. Overall, these results suggest that these two species differ strongly in many life-history traits as well as in the physiological control of their caste differentiation system. Possible origins and mechanisms of such interspecific variations are discussed, as well as their evolutionary and ecological consequences.

Variations in worker cuticular hydrocarbons and soldier isoprenoid defensive secretions within and among introduced and native populations of the subterranean termite, Reticulitermes flavipes.

In social insects, cuticular hydrocarbons (CHCs) play a central role in nestmate recognition. CHCs have proved to be useful for identifying species and differentiating populations. In combination with CHCs, isoprenoid soldier defensive secretions (SDSs) have been previously used in some termite species for chemotaxonomic analyses. This study compared the levels of chemical variation within and among introduced (French) and native (U.S.) populations of the subterranean termite, Reticulitermes flavipes. Worker CHCs and soldier SDSs from termites collected from colonies in nine populations in Florida, Louisiana, and France were analyzed. Discriminant analyses revealed that both localities and populations can be distinguished by using the variation in CHC profiles. Principal component analyses of CHC profiles as well as the calculation of two distance parameters (Nei and Euclidean) revealed remarkable chemical homogeneity within and among French populations. These analyses also showed that the CHC profiles of French populations were closer to termite populations from Louisiana than to those from Florida. Of the six distinct SDS chemotypes, one was common to populations in France and Louisiana. The possibility that populations in France originated from Louisiana, and the potential causes and consequences of chemical homogeneity within introduced populations are discussed.

Parasitoids modify their oviposition behavior according to the sexual origin of conspecific cuticular hydrocarbon traces.

Hydrocarbons play a crucial role in insect behavior in general and in sexual recognition in particular. Parasitoids often modify their oviposition behavior according to hydrocarbons left by conspecifics on the reproductive patch, such as oviposition markers left by females after oviposition, or cuticular hydrocarbon (CHC) traces left by individuals by walking or rubbing. This study determined whether Eupelmus vuilleti females are able to distinguish CHCs left by male or female conspecifics on seeds. The results show that the cuticular profile of E. vuilleti differs according to its gender, and that females are able to detect the sexual origin of these CHCs. Moreover, they adjust their oviposition behavior according to the nature of these traces. Although females lay fewer eggs on hosts when confronted with female CHCs, they lay more daughters when confronted with male CHCs, thus changing the sex ratio.