Mosaic Evolution of Molecular Pathways for Sex Pheromone Communication in a Butterfly.

Unraveling the origin of molecular pathways underlying the evolution of adaptive traits is essential for understanding how new lineages emerge, including the relative contribution of conserved ancestral traits and newly evolved derived traits. Here, we investigated the evolutionary divergence of sex pheromone communication from moths (mostly nocturnal) to butterflies (mostly diurnal) that occurred ~119 million years ago. In moths, it is the females that typically emit pheromones to attract male mates, but in butterflies males emit pheromones that are used by females for mate choice. The molecular bases of sex pheromone communication are well understood in moths, but they have remained relatively unexplored in butterflies. We used a combination of transcriptomics, real time qPCR, and phylogenetics to identify genes involved in the different steps (i.e., production, regulation, and reception) of sex pheromone communication of the butterfly Bicyclus anynana. Our results show that the biosynthesis and reception of sex pheromones relies both on moth-specific gene families (reductases) and on more ancestral insect gene families (desaturases, olfactory receptors, odorant binding proteins). Interestingly, B. anynana appears to use what was believed to be the moth-specific neuropeptide Pheromone Biosynthesis Activating Neuropeptide (PBAN) for regulating sex pheromone production. Altogether, our results suggest that a mosaic pattern best explains how sex pheromone communication evolved in butterflies, with some molecular components derived from moths, and others conserved from more ancient insect ancestors. This is the first large-scale investigation of the genetic pathways underlying sex pheromone communication in a butterfly.

Composition, Seasonal Variation, and Biological Activities of Lantana camara Essential Oils from Côte d'Ivoire.

This work aims to study the variations in the composition of Lantana camara leaf, flower, and stem essential oils over two years. L. camara organs were harvested in Bregbo (East Côte d'Ivoire) each month from June 2015 to June 2017. The essential oils were obtained by hydrodistillation and characterized by GC-MS and 13C NMR. Eighty-four compounds accounting for 84.4-99.1% of the essential oils have been identified. The essential oils hydrodistillated from L. camara are dominated by sesquiterpenes such as (E)-ß-caryophyllene and α-humulene, which were found in all samples. Some monoterpenes such as thymol, sabinene, and α-pinene were also present. Statistical analysis (principal component analysis and clustering) revealed a high variability in essential oil composition between the different organs and also within the studied periods, as the thymol proportion was higher during flowering and fruiting months. In addition, the stem, flower, and fruit essential oils were more concentrated in thymol than the leaf essential oils. The proportions of (E)-ß-caryophyllene and α-humulene were strictly inverted with the thymol proportion throughout the harvest period or vegetative cycle. The antioxidant, anti-inflammatory and insecticidal activities of leaves and flowers essential oils were also studied. Results showed that L. camara leaf and flower essential oils displayed high antioxidant, anti-inflammatory and insecticidal activities.

Generalized host-plant feeding can hide sterol-specialized foraging behaviors in bee-plant interactions.

Host-plant selection is a key factor driving the ecology and evolution of insects. While the majority of phytophagous insects is highly host specific, generalist behavior is quite widespread among bees and presumably involves physiological adaptations that remain largely unexplored. However, floral visitation patterns suggest that generalist bees do not forage randomly on all available resources. While resource availability and accessibility as well as nectar composition have been widely explored, pollen chemistry could also have an impact on the range of suitable host-plants. This study focuses on particular pollen nutrients that cannot be synthesized de novo by insects but are key compounds of cell membranes and the precursor for molting process: the sterols. We compared the sterol composition of pollen from the main host-plants of three generalist bees: Anthophora plumipes, Colletes cunicularius, and Osmia cornuta, as well as one specialist bee Andrena vaga. We also analyzed the sterols of their brood cell provisions, the tissues of larvae and nonemerged females to determine which sterols are used by the different species. Our results show that sterols are not used accordingly to foraging strategy: Both the specialist species A. vaga and the generalist species C. cunicularius might metabolize a rare C27 sterol, while the two generalist species A. plumipes and O. cornuta might rather use a very common C28 sterol. Our results suggest that shared sterolic compounds among plant species could facilitate the exploitation of multiple host-plants by A. plumipes and O. cornuta whereas the generalist C. cunicularius might be more constrained due to its physiological requirements of a more uncommon dietary sterol. Our findings suggest that a bee displaying a generalist foraging behavior may sometimes hide a sterol-specialized species. This evidence challenges the hypothesis that all generalist free-living bee species are all able to develop on a wide range of different pollen types.

Improving the Monitoring of the Walnut Husk Fly (Diptera: Tephritidae) Using Male-Produced Lactones.

It is important to monitor fruit flies (Diptera: Tephritidae) efficiently to implement sustainable means of control. Attractants are often used to increase the efficiency of sticky traps deployed in orchards to monitor Lepidopterans, but remains to be developed to monitor fruit flies. Rhagoletis completa Cresson (Diptera: Tephritidae) is an invasive species in the walnut orchards of Europe, and is commonly monitored with yellow sticky traps. In this study, we collected the volatile compounds released by male and female R. completa, and identified two lactones released exclusively by males. We then formulated both lactones in long-lasting volatile dispensers, and we quantified their release rate over a 26-d period. Finally, during the entire period when female flies are present in the field, we compared the efficiency of the conventional monitoring method using unbaited yellow sticky traps with yellow sticky traps associated with a dispenser releasing both male-produced lactones. These assays were conducted in 54 walnut orchards in France, in 2017. The number of fruit flies caught with sticky traps associated with lactones dispensers was increased by up to 10 times each week. Lactone-baited traps also allowed earlier detection in the season. These field results are promising for R. completa monitoring. A complete chiral identification of these lactones should be performed along with a clarification of their role in the sexual communication of R. completa.

Trace analysis of estrogenic compounds in surface and groundwater by ultra high performance liquid chromatography-tandem mass spectrometry as pyridine-3-sulfonyl derivatives.

Natural estrogens (estrone: E1, 17ß-estradiol: E2, estriol: E3) and synthetic 17α-ethynylestradiol (EE2) are reported as strong endocrine disruptors even at extremely low concentrations. Therefore, the watch list from the European Commission regarding emerging aquatic pollutants recommended maximum detection limits of 0.035 ng/L for EE2 and 0.4 ng/L for E1 and E2. In this study, a UHPLC-ESI-MS/MS method allowing quantification of E1, E2, E3 and EE2 in aqueous matrices was developed. The analytes were derivatized using pyridine-3-sulfonyl chloride and a broad range of product ions were generated and their specificity was assessed by analyzing both surface and groundwater. At least two product ions for each estrogenic compound were proved to be specific and hence suitable for quantification and confirmation. In complex aqueous matrices, analyte responses were particularly affected by ion suppression. This phenomenon was reduced by optimizing the clean-up and selecting a suitable stationary phase for the chromatographic separation. The limits of quantification assessed in surface water with the optimized method ranged from 0.098 ng/L (EE2) to 2.73 ng/L (E3).

Is non-host pollen suitable for generalist bumblebees?

Current evidence suggests that pollen is both chemically and structurally protected. Despite increasing interest in studying bee-flower networks, the constraints for bee development related to pollen nutritional content, toxicity and digestibility as well as their role in the shaping of bee-flower interactions have been poorly studied. In this study we combined bioassays of the generalist bee Bombus terrestris on pollen of Cirsium, Trifolium, Salix, and Cistus genera with an assessment of nutritional content, toxicity, and digestibility of pollen. Microcolonies showed significant differences in their development, non-host pollen of Cirsium being the most unfavorable. This pollen was characterized by the presence of quite rare δ7-sterols and a low digestibility. Cirsium consumption seemed increase syrup collection, which is probably related to a detoxification mixing behavior. These results strongly suggest that pollen traits may act as drivers of plant selection by bees and partly explain why Asteraceae pollen is rare in bee generalist diet.

Oviposition deterrent activity of basil plants and their essentials oils against Tuta absoluta (Lepidoptera: Gelechiidae).

The leafminer Tuta absoluta Meyrick (Lepidoptera: Gelechiidae) is one of the most important pests of tomato, reducing crop yields by up to 100% in greenhouses and fields, in several countries globally. Because synthetic insecticides lead to resistance and have adverse effects on natural enemies and the health of producers, alternative control methods are needed. In this study, we assessed the oviposition-deterring effect of basil plants, Ocimum gratissimum L. and O. basilicum L. (Lamiaceae), using dual-choice behavioural assays performed in flight tunnels. We found that both plants significantly reduced T. absoluta oviposition behaviour on a tomato plant located nearby. To evaluate the potential effect of basil volatile organic compounds, we formulated essential oils of both plant species in paraffin oil, and observed a similar oviposition-deterring effect. Gas chromatography analyses detected 18 constituents in these essential oils which the major constituents included thymol (33.3%), p-cymene (20.4%), γ-terpinene (16.9%), myrcene (3.9%) in O. gratissimum and estragol (73.8%), linalool (8.6%), ß-elemene (2.9%) and E-ß-ocimene (2.6%) in O. basilicum. Twenty and 33 compounds were identified of the volatiles collected on O. gratissimum and O. basilicum plants, respectively. The main components include the following: p-cymene (33.5%), γ-terpinene (23.6%), α-terpinene (7.2%), α-thujene (6.7%) and E-α-bergamotene (38.9%) in O. gratissimum, and methyl eugenol (26.1%), E-ß-ocimene (17.7%), and linalool (9.4%) in O. basilicum. Four compounds (α-pinene, ß-pinene, Myrcene, Limonene) were common in essential oils and plants. Our results suggest the valuable potential of basil and associated essential oils as a component of integrated management strategies against the tomato leafminer.

Could saponins be used to enhance bioremediation of polycyclic aromatic hydrocarbons in aged-contaminated soils?

Polycyclic aromatic hydrocarbons (PAH) are persistent organic compounds of major concern that tend to accumulate in the environment, threatening ecosystems and health. Brownfields represent an important tank for PAHs and require remediation. Researches to develop bioremediation and phytoremediation techniques are being conducted as alternatives to environmentally aggressive, expensive and often disruptive soil remediation strategies. The objectives of the present study were to investigate the potential of saponins (natural surfactants) as extracting agents and as bioremediation enhancers on an aged-contaminated soil. Two experiments were conducted on a brownfield soil containing 15 PAHs. In a first experiment, soil samples were extracted with saponins solutions (0; 1; 2; 4 and 8 g.L-1). In a second experiment conducted in microcosms (28 °C), soil samples were incubated for 14 or 28 days in presence of saponins (0; 2.5 and 5 mg g-1). CO2 emissions were monitored throughout the experiment. After the incubation, dehydrogenase activity was measured as an indicator of microbiological activity and residual PAHs were determined. In both experiments PAHs were determined using High-Performance Liquid Chromatography and Fluorimetric Detection. The 4 g.L-1 saponins solution extracted significantly more acenaphtene, fluorene, phenanthrene, anthracene, and pyrene than water. PAHs remediation was not enhanced in presence of saponins compared to control samples after 28 days. However CO2 emissions and dehydrogenase activities were significantly more important in presence of saponins, suggesting no toxic effect of these surfactants towards soil microbiota.

Identification of walnut husk (Juglans regia L.) volatiles and the behavioural response of the invasive Walnut Husk Fly, Rhagoletis completa Cresson.

BACKGROUND: Several European countries are important walnut (Juglans regia L.) producers. However, these countries must contend with the recent introduction of the Walnut Husk Fly, Rhagoletis completa Cresson (Diptera, Tephritidae), which is causing severe economic losses, especially in organic production. Because most Tephritid fruit flies use kairomones in their search for host plants, we hypothesise that this highly specialist species orients toward the volatile blend released by walnut husks. RESULTS: We collected, identified, and quantified the volatile organic chemicals (VOCs) released by walnut husks from the most commonly cultivated variety in France (Franquette). Then, the behavioural response of R. completa toward synthetic odour blends was recorded in dual choice assays conducted in net cages. A total of 26 VOCs were identified, with α-pinene, ß-pinene, trans-linalool, eugenol, and tetradecane representing the major constituents. In the dual choice assay, male and female R. completa were strongly attracted to synthetic blend that included most of the identified husk VOCs. CONCLUSION: When searching for a host plant, R. completa use host fruit kairomones. The potential of these semiochemicals in monitoring and management of this quarantine pest is discussed. © 2017 Society of Chemical Industry.

Minerals and Trace Elements Intakes and Food Consumption Patterns of Young Children Living in Rural Areas of Tibet Autonomous Region, P.R. China: A Cross-Sectional Survey.

BACKGROUND AND OBJECTIVES: Several studies revealed clinical signs of stunting and rickets among rural populations of Tibet Autonomous Region (T.A.R.), and especially amid children. Further, these populations are affected by a bone disease named Kashin-Beck disease (KBD). However, little is known about the dietary status of this population. This survey aimed to assess the usual intakes of young Tibetan children living in rural areas around Lhasa for energy, water, and ten minerals and trace elements (Na, K, Ca, P, Mg, Fe, Zn, Cu, Mn, and Se) involved in bone metabolism. DESIGN: A cross-sectional survey was designed. Totally, 250 pre-school children aged 3-5 years living in rural areas were enrolled. The 24-h food recall method was used to collect the intakes for two days, during two different seasons (September 2012 and April 2013). Because Tibetan foods are mainly derived from local agriculture and artisanal production, a combination of food composition tables was compiled, including specific and local food composition data. RESULTS: The Chinese dietary recommended intakes are not met for most of the elements investigated. Intake of sodium is much too high, while usual intakes are too low for K, Ca, Zn, Cu, and Se. Bioavailability of Ca, Fe, and Zn may be of concern due to the high phytic acid content in the diet. CONCLUSION: These nutrient imbalances may impact growth and bone metabolism of young Tibetan children. The advantages of the implementation of food diversification programs are discussed as well as the relevance of supplements distribution.

The importance of pollen chemistry in evolutionary host shifts of bees.

Although bee-plant associations are generally maintained through speciation processes, host shifts have occurred during evolution. Understanding shifts between both phylogenetically and morphologically unrelated plants (i.e., host-saltation) is especially important since they could have been key processes in the origin and radiation of bees. Probably far from being a random process, such host-saltation might be driven by hidden constraints associated with plant traits. We selected two clades of oligolectic bees (i.e., Colletes succinctus group and Melitta leporina group) foraging on co-flowering but unrelated host-plants to test this hypothesis. We analyzed floral scent, floral color and chemical composition of pollen from host and non-host plants of these two clades. We did not find evidence for host-plant evolution in the Melitta leporina group driven by one of the assayed floral traits. On the contrary, hosts of the C. succinctus group display similar primary nutritive content of pollen (i.e., amino acids and sterols) but not similar floral scent or color, suggesting that shared pollen chemistry probably mediates saltation in this clade. Our study revealed that constraints shaping floral associations are diverse and clearly depend on species life-history traits, but evidence suggests that pollen chemistry may act as a major floral filter and guide evolutionary host-shifts.

Looking for phosphate-accumulating bacteria in activated sludge processes: a multidisciplinary approach.

Over the past decades, an increasing need in renewable resources has progressively appeared. This trend concerns not only fossil fuels but also mineral resources. Wastewater and sewage sludge contain significant concentrations in phosphate and can be considered as a fertilizer source of the utmost importance. In wastewater treatment plants, the biological uptake of phosphate is performed by a specific microbiota: the phosphate-accumulating organisms. These microorganisms are recovered in sewage sludge. Here, we aimed to investigate the occurrence of phosphate accumulators in four wastewater treatment plants. A 16S metagenetic analysis identified the main bacterial phyla extracted from the aerobic treatment: α-Proteobacteria, ß-Proteobacteria, and Sphingobacteria. An enrichment stage was performed to stimulate the specific growth of phosphate-accumulating bacteria in an acetate medium. An analysis of metabolic activities of sulfur and phosphorus highlighted strong modifications related to phosphorus and much less distinguishable effects with sulfur. A solid acetate medium containing 5-Br-4-Cl-3-indolyl phosphate was used to select potential phosphate-accumulating bacteria from the enriched consortia. The positive strains have been found to belong in the genera Acinetobacter, Corynebacterium, and Pseudomonas. Finally, electron microscopy was applied to the strains and allowed to confirm the presence of polyphosphate granules. Some of these bacteria contained granules the size of which exceeded 100 nm.

Elevated Carbon Dioxide Concentration Reduces Alarm Signaling in Aphids.

Insects often rely on olfaction to communicate with conspecifics. While the chemical language of insects has been deciphered in recent decades, few studies have assessed how changes in atmospheric greenhouse gas concentrations might impact pheromonal communication in insects. Here, we hypothesize that changes in the concentration of atmospheric carbon dioxide affect the whole dynamics of alarm signaling in aphids, including: (1) the production of the active compound (E)-ß-farnesene (Eßf), (2) emission behavior when under attack, (3) perception by the olfactory apparatus, and (4) the escape response. We reared two strains of the pea aphid, Acyrthosiphon pisum, under ambient and elevated CO2 concentrations over several generations. We found that an increase in CO2 concentration reduced the production (i.e., individual content) and emission (released under predation events) of Eßf. While no difference in Eßf neuronal perception was observed, we found that an increase in CO2 strongly reduced the escape behavior expressed by an aphid colony following exposure to natural doses of alarm pheromone. In conclusion, our results confirm that changes to greenhouse gases impact chemical communication in the pea aphid, and could potentially have a cascade effect on interactions with higher trophic levels.

Impact of pollen resources drift on common bumblebees in NW Europe.

Several bee species are experiencing significant population declines. As bees exclusively rely on pollen for development and survival, such declines could be partly related to changes in their host plant abundance and quality. Here, we investigate whether generalist bumblebee species, with stable population trends over the past years, adapted their diets in response to changes in the distribution and chemical quality of their pollen resources. We selected five common species of bumblebee in NW Europe for which we had a precise description of their pollen diet through two time periods ('prior to 1950' and '2004-2005'). For each species, we assessed whether the shift in their pollen diet was related with the changes in the suitable area of their pollen resources. Concurrently, we evaluated whether the chemical composition of pollen resources changed over time and experimentally tested the impact of new major pollen species on the development of B. terrestris microcolonies. Only one species (i.e. B. lapidarius) significantly included more pollen from resources whose suitable area expanded. This opportunist pattern could partly explain the expansion of B. lapidarius in Europe. Regarding the temporal variation in the chemical composition of the pollen diet, total and essential amino acid contents did not differ significantly between the two time periods while we found significant differences among plant species. This result is driven by the great diversity of resources used by bumblebee species in both periods. Our bioassay revealed that the shift to new major pollen resources allowed microcolonies to develop, bringing new evidence on the opportunist feature of bumblebee in their diets. Overall, this study shows that the response to pollen resource drift varies among closely related pollinators, and a species-rich plant community ensures generalist species to select a nutrient-rich pollen diet.

Bacteria may contribute to distant species recognition in ant-aphid mutualistic relationships.

Mutualistic interactions between ant and aphid species have been the subject of considerable historical and contemporary investigations, the primary benefits being cleaning and protection for the aphids and carbohydrate-rich honeydew for the ants. Questions remained, however, as to the volatile semiochemical factor influencing this relationship. A recent study highlighted the role of bacterial honeydew volatile compounds in ant attraction. Here, ant's ability to distantly discriminate 2 aphid species was investigated based on bacterial honeydew semiochemicals emissions using a two-way olfactometer. Both the mutualistic aphid Aphis fabae L. and the nonmyrmecophilous aphid Acyrthosiphon pisum Harris were found to be attractive for the ant Lasius niger L. The level of attraction was similar in both assays (control vs. one of the aphid species). However, when given a choice between these 2 aphid species, ants showed a significant preference for Aphis fabae. Honeydew volatiles, mostly from bacterial origins, are known to be a key element in ant attraction. Using the same olfactometry protocol, the relative attractiveness of volatiles emitted by honeydews collected from each aphid species and by bacteria isolated from each honeydew was investigated. Again, ants significantly preferred volatiles released by Aphis fabae honeydew and bacteria. This information suggests that microbial honeydew volatiles enable ants to distantly discriminate aphid species. These results strengthen the interest of studying the occurrence and potential impact of microorganisms in insect symbioses.

Chemical Composition and Acaricidal Activity of Thymus algeriensis Essential Oil against Varroa destructor.

The aim of the present study was to determine the chemical composition and evaluate the acaricidal activity of Thymus algeriensis essential oil (TAEO) against Varroa destructor. This ectoparasitic mite is a pest of the honey bee Apis mellifera. The essential oil from the, aerial parts of T. algeriensis, obtained by hydrodistillation, was obtained in a yield of 2.8± 0.2%, w/w. The TAEO was analyzed by GC and GC/MS. Thirty-four compounds were identified, representing 99.3% of the oil. The main constituents were carvacrol (48.4%), γ-terpinene (14.9%), p-cymene (14.7%), and thymol (5.6%). Four lots 'were constituted at the level of an apiary in order to study the dynamics of the Varroa destructor and its host, Apis mellifera. After diagnosis by the.biological method "install of diapers", the lots were treated at different doses of TAEO (0.1, 0.3 and 0.5%). TAEO was sprayed on top of the hives. The results show that TAEO at 0.5% resulted in a decrease in the rate of infestation of Varroa destructor, causing a mortality rate of 32.6% without negative effect on the nesting of the queen. The essential oil of T. algeriensis could be used as a bioacaricidal agent.

Analytical methods used to quantify isoflavones in cow's milk: a review.

This paper provides an update and comprehensive review of the analytical methods used for quantifying isoflavones and their metabolites in cow's milk. Isoflavones are secondary plant metabolites that are similar to 17 ß-estradiol in chemical structure. They form one of the most common categories of phytoestrogens. Numerous health benefits have been attributed to isoflavones, but many of these compounds are also considered to be endocrine disruptors, with adverse effects on health. These contradictory trends offer an attractive prospect for future research, and therefore, sensitive and reliable analytical methods are required to clarify various issues about isoflavones. For this review, a structured methodology was used to select 26 relevant articles published between 2005 and 2015 from the Scopus and CAB Abstract databases. The review discusses individual steps of the analytical procedures described in these articles, including sample preparation, instrumental analysis and validation. The most commonly used analytical procedure is sample preparation involving liquid-liquid extraction and an enzymatic hydrolysis step followed by liquid chromatography with mass spectrometry analysis. Currently, however, there is no standardized procedure for the sample preparation and analysis of isoflavones in milk.

Effects of thirty elements on bone metabolism.

The human skeleton, made of 206 bones, plays vital roles including supporting the body, protecting organs, enabling movement, and storing minerals. Bones are made of organic structures, intimately connected with an inorganic matrix produced by bone cells. Many elements are ubiquitous in our environment, and many impact bone metabolism. Most elements have antagonistic actions depending on concentration. Indeed, some elements are essential, others are deleterious, and many can be both. Several pathways mediate effects of element deficiencies or excesses on bone metabolism. This paper aims to identify all elements that impact bone health and explore the mechanisms by which they act. To date, this is the first time that the effects of thirty minerals on bone metabolism have been summarized.

Do aphids actively search for ant partners?

The aphid-ant mutualistic relationships are not necessarily obligate for neither partners but evidence is that such interactions provide them strong advantages in terms of global fitness. While it is largely assumed that ants actively search for their mutualistic partners namely using volatile cues; whether winged aphids (i.e., aphids' most mobile form) are able to select ant-frequented areas had not been investigated so far. Ant-frequented sites would indeed offer several advantages for these aphids including a lower predation pressure through ant presence and enhanced chances of establishing mutuaslistic interactions with neighbor ant colonies. In the field, aphid colonies are often observed in higher densities around ant nests, which is probably linked to a better survival ensured by ants' services. Nevertheless, this could also result from a preferential establishment of winged aphids in ant-frequented areas. We tested this last hypothesis through different ethological assays and show that the facultative myrmecophilous black bean aphid, Aphis fabae L., does not orientate its search for a host plant preferentially toward ant-frequented plants. However, our results suggest that ants reduce the number of winged aphids leaving the newly colonized plant. Thus, ants involved in facultative myrmecophilous interactions with aphids appear to contribute to structure aphid populations in the field by ensuring a better establishment and survival of newly established colonies rather than by inducing a deliberate plant selection by aphid partners based on the proximity of ant colonies.

First evidence of a volatile sex pheromone in lady beetles.

To date, volatile sex pheromones have not been identified in the Coccinellidae family; yet, various studies have suggested that such semiochemicals exist. Here, we collected volatile chemicals released by virgin females of the multicolored Asian lady beetle, Harmonia axyridis (Pallas), which were either allowed or not allowed to feed on aphids. Virgin females in the presence of aphids, exhibited "calling behavior", which is commonly associated with the emission of a sex pheromone in several Coleoptera species. These calling females were found to release a blend of volatile compounds that is involved in the remote attraction (i.e., from a distance) of males. Gas Chromatography-Mass Spectrometry (GC-MS) analyses revealed that (-)-ß-caryophyllene was the major constituent of the volatile blend (ranging from 80 to 86%), with four other chemical components also being present; ß-elemene, methyl-eugenol, α-humulene, and α-bulnesene. In a second set of experiments, the emission of the five constituents identified from the blend was quantified daily over a 9-day period after exposure to aphids. We found that the quantity of all five chemicals significantly increased across the experimental period. Finally, we evaluated the activity of a synthetic blend of these chemicals by performing bioassays which demonstrated the same attractive effect in males only. The results confirm that female H. axyridis produce a volatile sex pheromone. These findings have potential in the development of more specific and efficient biological pest-control management methods aimed at manipulating the behavior of this invasive lady beetle.