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This work focuses on the profiling of semiochemicals (SCS) from Callosobruchus maculatus , Sitophilus oryzae and Tribolium castaneum by headspace solid-phase microextraction (HS-SPME) coupled with gas chromatography-mass spectrometry (GC-MS) technique. Totally, 6, 9 and 8 volatile compounds (VCS) were detected from C. maculatus , S. oryzae and T. castaneum , respectively. As a result of pherobase analysis and preference bioassay, stearic acid ( C. maculatus ); nonanal, lauric acid and stearic acid ( S. oryzae ); stearic acid ( T. castaneum ) were identified as new SCS that could be useful for integrated pest management (IPM) practices.

Geosmin, a ubiquitous volatile sesquiterpenoid of microbiological origin, is causative for deteriorating the quality of many foods, beverages, and drinking water, by eliciting an undesirable "earthy/musty" off-flavor. Moreover, and across species from worm to human, geosmin is a volatile, chemosensory trigger of both avoidance and attraction behaviors, suggesting its role as semiochemical. Volatiles typically are detected by chemosensory receptors of the nose, which have evolved to best detect ecologically relevant food-related odorants and semiochemicals. An insect receptor for geosmin was recently identified in flies. A human geosmin-selective receptor, however, has been elusive. Here, we report on the identification and characterization of a human odorant receptor for geosmin, with its function being conserved in orthologs across six mammalian species. Notably, the receptor from the desert-dwelling kangaroo rat showed a more than 100-fold higher sensitivity compared to its human ortholog and detected geosmin at low nmol/L concentrations in extracts from geosmin-producing actinomycetes.

Background and Aims: Cress seeds release allelochemicals that over-stimulate the elongation of hypocotyls of neighbouring (potentially competing) seedlings and inhibit their root growth. The hypocotyl promoter is potassium, but the root inhibitor was unidentified; its nature is investigated here., Methods: Low-molecular-weight cress-seed exudate (LCSE) from imbibed Lepidium sativum seeds was fractionated by phase partitioning, paper chromatography, high-voltage electrophoresis and gel-permeation chromatography (on Bio-Gel P-2). Fractions, compared with pure potassium salts, were bioassayed for effects on Amaranthus caudatus seedling growth in the dark for 4 days., Key Results: The LCSE robustly promoted amaranth hypocotyl elongation and inhibited root growth. The hypocotyl inhibitor was non-volatile, hot acid stable, hydrophilic and resistant to incineration, as expected for K+. The root inhibitor(s) had similar properties but were organic (activity lost on incineration). The root inhibitor(s) remained in the aqueous phase (at pH 2.0, 6.5 and 9.0) when partitioned against butan-1-ol or toluene, and were thus hydrophilic. Activity was diminished after electrophoresis, but the remaining root inhibitors were neutral. They became undetectable after paper chromatography; therefore, they probably comprised multiple compounds, which separated from each other, in part, during fractionation. On gel-permeation chromatography, the root inhibitor co-eluted with hexoses., Conclusions: Cress-seed allelochemicals inhibiting root growth are different from the agent (K+) that over-stimulates hypocotyl elongation and the former probably comprise a mixture of small, non-volatile, hydrophilic, organic substances. Abundant components identified chromatographically and by electrophoresis in cress-seed exudate fitting this description include glucose, fructose, sucrose and galacturonic acid. However, none of these sugars co-chromatographed and co-electrophoresed with the root-inhibitory principle of LCSE, and none of them (in pure form at naturally occurring concentrations) inhibited root growth. We conclude that the root-inhibiting allelochemicals of cress-seed exudate remain unidentified., (© The Author(s) 2023. Published by Oxford University Press on behalf of the Annals of Botany Company. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

In this work, we identified the trail pheromone of the ant Crematogaster scutellaris. We combined gas chromatography-mass spectrometry analysis of extracts from the hind tibia, the location of the respective glands, with automated trail following assays. The study found tridecan-2-ol to be the strongest discriminator between hind tibia and other body part extracts. Tridecan-2-ol elicited trail-following behaviour at concentrations of 1 ng/µL. A separation of the enantiomers showed responses to (R)-tridecan-2-ol already at 0.001 ng/µL and only at a 1000-fold higher concentration for (S)-tridecan-2-ol, suggesting that only the R enantiomer is used by C. scutellaris in its natural environment. We also found strong behavioural responses to 2-dodecanol, a substance that was not detectable in the hind tibia extract of C. scutellaris, but which has been reported to be the trail pheromone of the related species C. castanea. We discuss the contribution of these results to the 'dissection and reconstruction' of strategies and mechanisms underlying the social organization of ants., (© 2024. The Author(s).)

Competition for soil nutrients and water with other plants foster competition within the biosphere for access to these limited resources. The roots for the common grain sorghum produce multiple small molecules that are released via root exudates into the soil to compete with other plants. Sorgoleone is one such compound, which suppresses weed growth near sorghum by acting as a quinone analog and interferes with photosynthesis. Since sorghum also grows photosynthetically, and may be susceptible to sorgoleone action if present in tissues above ground, it is essential to exude sorgoleone efficiently. However, since the P450 enzymes that synthesize sorgoleone are intracellular, the release mechanism for sorgoleone remain unclear. In this study, we conducted an in silico assessment for sorgoleone and its precursors to passively permeate biological membranes. To facilitate accurate simulation, CHARMM parameters were newly optimized for sorgoleone and its precursors. These parameters were used to conduct 1 μs of unbiased molecular dynamics simulations to compare the permeability of sorgoleone with its precursors molecules. We find that interleaflet transfer is maximized for sorgoleone, suggesting that the precursor molecules may remain in the same leaflet for access by biosynthetic P450 enzymes. Since no sorgoleone was extracted during unbiased simulations, we compute a permeability coefficient using the inhomogeneous solubility diffusion model. The requisite free energy and diffusivity profiles for sorgoleone through a sorghum membrane model were determined through Replica Exchange Umbrella Sampling (REUS) simulations. The REUS calculations highlight that any soluble sorgoleone would quickly insert into a lipid bilayer, and would readily transit. When sorgoleone forms aggregates in root exudate as indicated by our equilibrium simulations, aggregate formation would lower the effective concentration in aqueous solution, creating a concentration gradient that would facilitate passive transport. This suggests that sorgoleone synthesis occurs within sorghum root cells and that sorgoleone is exuded by permeating through the cell membrane without the need for a transport protein once the extracellular sorgoleone aggregate is formed., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

Background: Plants growing in proximity to other plants are exposed to a variety of metabolites that these neighbors release into the environment. Some species produce allelochemicals to inhibit growth of neighboring plants, which in turn have evolved ways to detoxify these compounds., Results: In order to understand how the allelochemical-receiving target plants respond to chemically diverse compounds, we performed whole-genome transcriptome analysis of Arabidopsis thaliana exposed to either the benzoxazinoid derivative 2-amino- 3H-phenoxazin-3-one (APO) or momilactone B. These two allelochemicals belong to two very different compound classes, benzoxazinoids and diterpenes, respectively, produced by different Poaceae crop species., Conclusions: Despite their distinct chemical nature, we observed similar molecular responses of A. thaliana to these allelochemicals. In particular, many of the same or closely related genes belonging to the three-phase detoxification pathway were upregulated in both treatments. Further, we observed an overlap between genes upregulated by allelochemicals and those involved in herbicide detoxification. Our findings highlight the overlap in the transcriptional response of a target plant to natural and synthetic phytotoxic compounds and illustrate how herbicide resistance could arise via pathways involved in plant-plant interaction., (© 2022. The Author(s).)

Mate choice informed by the immune genes of the major histocompatibility complex (MHC) may provide fitness benefits including offspring with increased immunocompetence. Olfactory cues are considered the primary mechanism organisms use to evaluate the MHC of potential mates, yet this idea has received limited attention in birds. Motivated by a finding of MHC-dependent mate choice in the Leach's storm-petrel ( Oceanodroma leucorhoa ), we examined whether the chemical profiles of this highly scented seabird contain information about MHC genes. Whereas previous studies in birds examined non-volatile compounds, we used gas chromatography-mass spectrometry to measure the volatile compounds emitted from feathers that potentially serve as olfactory infochemicals about MHC and coupled this with locus-specific genotyping of MHC IIB genes. We found that feather chemicals reflected individual MHC diversity through interactions with sex and breeding status. Furthermore, similarity in MHC genotype was correlated with similarity in chemical profiles within female-female and male-female dyads. We provide the first evidence that volatile chemicals from bird feathers can encode information about the MHC. Our findings suggest that olfaction likely aids MHC-based mate choice in this species and highlight a role for chemicals in mediating genetic mate choice in birds where this mode of communication has been largely overlooked.

Background: Eurasian spruce bark beetles (Ips typographus) use both attractant and anti-attractant semiochemicals to find suitable mature host trees. Trans-4-thujanol is abundant in young, unsuitable spruce trees. Electrophysiological studies have demonstrated its high activity levels, but field data are lacking., Results: Enantioselective GC-MS analysis showed that only (1R,4S)-(+)-trans-4-thujanol was present in Norway spruce bark volatiles. In a factorial design field-trapping experiment, trans-4-thujanol alone was not attractive to Ips typographus. Traps baited with I. typographus' aggregation pheromone and trans-4-thujanol or the known anti-attractant 1,8-cineole caught fewer beetles than those baited with the aggregation pheromone alone. Catches for trans-4-thujanol and 1,8-cineole were dose-dependent. Intermediate doses of trans-4-thujanol and 1,8-cineole had a similar effect. Surprisingly, in contrast to 1,8-cineole and other known Ips anti-attractants, which all inhibit males more strongly than females, the addition of trans-4-thujanol to the aggregation pheromone reduced the attraction of females more., Conclusion: The Norway spruce volatile (+)-trans-4-thujanol is a novel I. typographus anti-attractant with potency comparable to the known anti-attractants 1,8-cineole and verbenone, and is more effective for females than for males. Incorporating (+)-trans-4-thujanol into anti-attractant lures could improve protection of trees from mass attack by I. typographus., (© 2022 Society of Chemical Industry.)

Background: The flowers of some species of orchids produce nectar as a reward for pollination, the process of transferring pollen from flower to flower. Epipactis albensis is an obligatory autogamous species, does not require the presence of insects for pollination, nevertheless, it has not lost the ability to produce nectar, the chemical composition of which we examined by gas chromatography-mass spectrometry (GC-MS) method for identification of potential insect attractants., Results: During five years of field research, we did not observe any true pollinating insects visiting the flowers of this species, only accidental insects as ants and aphids. As a result of our studies, we find that this self-pollinating orchid produces in nectar inter alia aliphatic saturated and unsaturated aldehydes such as nonanal (pelargonal) and 2-pentenal as well as aromatic ones (i.e., syringaldehyde, hyacinthin). The nectar is low in alkenes, which may explain the absence of pollinating insects. Moreover, vanillin and eugenol derivatives, well-known as important scent compounds were also identified, but the list of chemical compounds is much poorer compared with a closely related species, insect-pollinating E. helleborine., Conclusion: Autogamy is a reproductive mechanism employed by many flowering plants, including the orchid genus Epipactis, as an adaptation to growing in habitats where pollinating insects are rarely observed due to the lack of nectar-producing plants they feed on. The production of numerous chemical attractants by self-pollinated E. albensis confirms the evolutionary secondary process, i.e., transition from ancestral insect-pollinating species to obligatory autogamous., (© 2022. The Author(s).)

Insect-pollinated plants often release complex mixtures of floral scents to attract their pollinators. Yet scent compounds eliciting physiological or behavioural responses in pollinators have only been identified in few plant species. The sapromyiophilous aroid Arum maculatum releases a highly diverse dung-like scent with overall more than 300 different compounds recorded so far to attract its psychodid and other fly pollinators. The volatiles' role in pollinator attraction is mostly unknown. To identify potential behaviourally active compounds, we recorded electroantennographic responses of four Psychodidae and one Sphaeroceridae species to (1) inflorescence scents of A. maculatum and (2) the scents released by cow dung, likely imitated by the plant species. Here we show that these flies are sensitive to 78 floral volatiles of various chemical classes, 18 of which were also found in cow dung. Our study, which for the first time determined physiologically active compounds in the antennae of Psychoda spp. and Sphaeroceridae, identified various volatiles not known to be biologically active in any floral visitors so far. The obtained results help deciphering the chemical basis that enables A. maculatum and other plants, pollinated by psychodids and sphaerocerids, to attract and deceive their pollinators., (© 2022. The Author(s).)

Vespa velutina has been rapidly expanding throughout Galicia since 2012. It is causing human health risks and well-known losses in the beekeeping sector. Control methods are scarce, unspecific, and ineffective. Semiochemicals are insect-derived chemicals that play a role in communication and they could be used an integrated pest management tool alternative to conventional pesticides. A previous determination of the organic chemical profile should be the first step in the study of these semiochemicals. HS-SPME in living individuals and the sting apparatus extraction followed by GC-MS spectrometry were combined to extract a possible profile of these compounds in 43 hornets from Galicia. The identified compounds were hydrocarbons, ketones, terpenes, and fatty acid, and fatty acid esters. Nonanal aldehyde appeared in important concentrations in living individuals. While pentadecane, 8-hexyl- and ethyl oleate were mainly extracted from the venom apparatus. Ketones 2-nonanone, 2-undecanone and 7-nonen-2-one, 4,8-dimethyl- were identified by both procedures, as was 1,7-Nonadiene, 4,8-dimethyl-. Some compounds were detected for the first time in V. velutina such as naphthalene, 1,6-dimethyl-4-(1-methylethyl). The chemical profile by caste was also characterized.

Loxosceles reclusa, or brown recluse spider, is a harmful household spider whose habitat extends throughout the Midwest in the USA and other regions in the world. The pheromones and other biomolecules that facilitate signaling for brown recluses and other spider species are poorly understood. A rapid and sensitive method is needed to analyze airborne spider signaling biomolecules to better understand the structure and function of these biochemicals in order to control the population of the spiders. In this study, we developed a novel headspace solid-phase microextraction (HS-SPME)-GC/MS method to analyze potential pheromones and biomolecules emitted by the brown recluse spider. The method is highly selective and sensitive for biomolecule identification and quantification from a single live spider. Using this novel non-destructive HS-SPME-GC/MS technique, we identified 11 airborne biomolecules, including 4-methylquinazoline, dimethyl sulfone, 2-methylpropanoic acid, butanoic acid, hexanal, 3-methylbutanoic acid, 2-methylbutanoic acid, 2,4-dimethylbenzaldehyde, 2-phenoxyethanol, and citral (contains both isomers of neral and geranial). Some of these airborne biomolecules were also reported as semiochemicals associated with biological functions of other spiders and insects. The method was also applied to study the airborne biochemicals of Plectreurys tristis, another primitive hunting spider with a poor web, enabling quantitation of the same compounds and demonstrating a difference in signaling molecule concentrations between the two species. This method has potential application in the study of pheromones and biological signaling in other species, which allows for the possibility of utilizing attractant or deterrent functions to limit household populations of harmful species., (© 2021. Springer-Verlag GmbH Germany, part of Springer Nature.)

Timely and accurate diagnostics are essential to fight the COVID-19 pandemic, but no test satisfies both conditions. Dogs can scent-identify the unique odors of volatile organic compounds generated during infection by interrogating specimens or, ideally, the body of a patient. After training 6 dogs to detect SARS-CoV-2 by scent in human respiratory secretions (in vitro diagnosis), we retrained 5 of them to search and find the infection by scenting the patient directly (in vivo screening). Then, efficacy trials were designed to compare the diagnostic performance of the dogs against that of the rRT-PCR in 848 human subjects: 269 hospitalized patients (COVID-19 prevalence 30.1%), 259 hospital staff (prevalence 2.7%), and 320 government employees (prevalence 1.25%). The limit of detection in vitro was lower than 10-12 copies ssRNA/mL. During in vivo efficacy experiments, our 5 dogs detected 92 COVID-19 positive patients among the 848 study subjects. The alert (lying down) was immediate, with 95.2% accuracy and high sensitivity (95.9%; 95% C.I. 93.6-97.4), specificity (95.1%; 94.4-95.8), positive predictive value (69.7%; 65.9-73.2), and negative predictive value (99.5%; 99.2-99.7) in relation to rRT-PCR. Seventy-five days after finishing in vivo efficacy experiments, a real-life study (in vivo effectiveness) was executed among the riders of the Metro System of Medellin, deploying the human-canine teams without previous training or announcement. Three dogs were used to examine the scent of 550 volunteers who agreed to participate, both in test with canines and in rRT-PCR testing. Negative predictive value remained at 99.0% (95% C.I. 98.3-99.4), but positive predictive value dropped to 28.2% (95% C.I. 21.1-36.7). Canine scent-detection in vivo is a highly accurate screening test for COVID-19, and it detects more than 99% of infected individuals independent of key variables, such as disease prevalence, time post-exposure, or presence of symptoms. Additional training is required to teach the dogs to ignore odoriferous contamination under real-life conditions., Competing Interests: The authors have declared that no competing interests exist.

Memorizing pheromonal locations is critical for many mammalian species as it involves finding mates and avoiding competitors. In rodents, pheromonal information is perceived by the main and accessory olfactory systems. However, the role of somatosensation in context-dependent learning and memorizing of pheromone locations remains unexplored. We addressed this problem by training female mice on a multimodal task to locate pheromones by sampling volatiles emanating from male urine through the orifices of varying dimensions or shapes that are sensed by their vibrissae. In this novel pheromone location assay, female mice' preference toward male urine scent decayed over time when they were permitted to explore pheromones vs neutral stimuli, water. On training them for the associations involving olfactory and whisker systems, it was established that they were able to memorize the location of opposite sex pheromones, when tested 15 days later. This memory was not formed either when the somatosensory inputs through whisker pad were blocked or when the pheromonal cues were replaced with that of same sex. The association between olfactory and somatosensory systems was further confirmed by the enhanced expression of the activity-regulated cytoskeleton protein. Furthermore, the activation of main olfactory bulb circuitry by pheromone volatiles did not cause any modulation in learning and memorizing non-pheromonal volatiles. Our study thus provides the evidence for associations formed between different sensory modalities facilitating the long-term memory formation relevant to social and reproductive behaviors., (© 2021 The Authors. The FASEB Journal published by Wiley Periodicals LLC on behalf of Federation of American Societies for Experimental Biology.)

Aphids are destructive pests, and alarm pheromones play a key role in their chemical ecology. Here, we conducted a detailed analysis of terpenoids in the vetch aphid, Megoura viciae, and its host plant Pisum sativum using gas chromatography/mass spectrometry. Four major components, (-)-β-pinene (49.74%), (E)-β-farnesene (32.64%), (-)-α-pinene (9.42%) and ( +)-limonene (5.24%), along with trace amounts of ( +)-sabinene, camphene and α-terpineol) (3.14%) were found in the aphid. In contrast, few terpenoids were found in the host plant, consisting mainly of squalene (66.13%) and its analog 2,3-epoxysqualene (31.59%). Quantitative analysis of the four major terpenes in different developmental stages of the aphid showed that amounts of the monoterpenes increased with increasing stage, while the sesquiterpene amount peaked in the 3 rd instar. (-)-β-Pinene was the most abundant terpene at all developmental stages. Behavioral assays using a three-compartment olfactometer revealed that the repellency of single compounds varied in a concentration-dependent manner, but two mixtures [(-)-α-pinene: (-)-β-pinene: (E)-β-farnesene: ( +)-limonene = 1:44.4:6.5:2.2 or 1:18.4:1.3:0.8], were repellent at all concentrations tested. Our results suggest that (-)-α-pinene and (-)-β-pinene are the major active components of the alarm pheromone of M. viciae, but that mixtures play a key role in the alarm response. Our study contributes to the understanding of the chemical ecology of aphids and may help design new control strategies against this aphid pest., (© 2021. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

Pheromones are biologically important in fruit fly mating systems, and also have potential applications as attractants or mating disrupters for pest management. Bactrocera kraussi (Hardy) (Diptera: Tephritidae) is a polyphagous pest fruit fly for which the chemical profile of rectal glands is available for males but not for females. There have been no studies of the volatile emissions of either sex or of electrophysiological responses to these compounds. The present study (i) establishes the chemical profiles of rectal gland contents and volatiles emitted by both sexes of B. kraussi by gas chromatography-mass spectrometry (GC-MS) and (ii) evaluates the detection of the identified compounds by gas chromatography-electroantennogram detection (GC-EAD) and -electropalpogram detection (GC-EPD). Sixteen compounds are identified in the rectal glands of male B. kraussi and 29 compounds are identified in the rectal glands of females. Of these compounds, 5 were detected in the headspace of males and 13 were detected in the headspace of females. GC-EPD assays recorded strong signals in both sexes against ( E , E )-2,8-dimethyl-1,7-dioxaspiro[5.5]undecane, 2-ethyl-7-mehtyl-1,6-dioxaspiro[4.5]decane isomer 2, ( E , Z )/( Z , E )-2,8-dimethyl-1,7-dioxaspiro[5.5]undecane, and ( Z , Z )-2,8-dimethyl-1,7-dioxaspiro[5.5]undecane. Male antennae responded to ( E , E )-2,8-dimethyl-1,7-dioxaspiro[5.5]undecane, 2-methyl-6-pentyl-3,4-dihydro-2 H -pyran, 6-hexyl-2-methyl-3,4-dihydro-2 H -pyran, 6-oxononan-1-ol, ethyl dodecanoate, ethyl tetradecanoate and ethyl ( Z )-hexadec-9-enoate, whereas female antennae responded to ( E , E )-2,8-dimethyl-1,7-dioxaspiro[5.5]undecane and 2-methyl-6-pentyl-3,4-dihydro-2 H -pyran only. These compounds are candidates as pheromones mediating sexual interactions in B. kraussi .

The polyphagous invasive brown marmorated stink bug, Halyomorpha halys, reportedly discriminates among phenological stages of host plants. To determine whether olfaction is involved in host plant stage discrimination, we selected (dwarf) sunflower, Helianthus annuus, as a model host plant species. When adult females of a still-air laboratory experiment were offered a choice of four potted sunflowers at distinct phenological stages (vegetative, pre-bloom, bloom, seeding), most females settled onto blooming plants but oviposited evenly on plants of all four stages. In moving-air two-choice olfactometer experiments, we then tested each plant stage versus filtered air and versus one another, for attraction of H. halys females. Blooming sunflowers performed best overall, but no one plant stage was most attractive in all experiments. Capturing and analyzing (by GC-MS) the headspace odorants of each plant stage revealed a marked increase of odorant abundance (e.g., monoterpenes) as plants transitioned from pre-bloom to bloom. Analyzing the headspace odorant blend of blooming sunflower by gas chromatographic-electroantennographic detection (GC-EAD) revealed 13 odorants that consistently elicited responses from female H. halys antennae. An 11-component synthetic blend of these odorants attracted H. halys females in laboratory olfactometer experiments. Furthermore, in field settings, the synthetic blend enhanced the attractiveness of synthetic H. halys pheromone as a trap lure, particularly in spring (April to mid-June). A simpler yet fully effective sunflower semiochemical blend could be developed and coupled with synthetic H. halys aggregation pheromones to improve monitoring efforts or could improve the efficacy of modified attract-and-kill control tactics for H. halys., (© 2021. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

Molecular recognition is a fundamental principle in biological systems. The olfactory detection of both food and predators via ecological relevant odorant cues are abilities of eminent evolutionary significance for many species. Pyrazines are such volatile cues, some of which act as both human-centered key food odorants (KFOs) and semiochemicals. A pyrazine-selective odorant receptor has been elusive. Here we screened 2,3,5-trimethylpyrazine, a KFO and semiochemical, and 2,5-dihydro-2,4,5-trimethylthiazoline, an innate fear-associated non-KFO, against 616 human odorant receptor variants, in a cell-based luminescence assay. OR5K1 emerged as sole responding receptor. Tested against a comprehensive collection of 178 KFOs, we newly identified 18 pyrazines and (2R/2S)-4-methoxy-2,5-dimethylfuran-3(2H)-one as agonists. Notably, OR5K1 orthologs in mouse and domesticated species displayed a human-like, potency-ranked activation pattern of pyrazines, suggesting a domestication-led co-evolution of OR5K1 and its orthologs. In summary, OR5K1 is a specialized olfactory receptor across mammals for the detection of pyrazine-based key food odors and semiochemicals., (© 2021 The Authors. The FASEB Journal published by Wiley Periodicals LLC on behalf of Federation of American Societies for Experimental Biology.)

The Annonaceae fruits weevil ( Optatus palmaris ) causes high losses to the soursop production in Mexico. Damage occurs when larvae and adults feed on the fruits; however, there is limited research about control strategies against this pest. However, pheromones provide a high potential management scheme for this curculio. Thus, this research characterized the behavior and volatile production of O. palmaris in response to their feeding habits. Olfactometry assays established preference by weevils to volatiles produced by feeding males and soursop. The behavior observed suggests the presence of an aggregation pheromone and a kairomone. Subsequently, insect volatiles sampled by solid-phase microextraction and dynamic headspace detected a unique compound on feeding males increased especially when feeding. Feeding-starvation experiments showed an averaged fifteen-fold increase in the concentration of a monoterpenoid on males feeding on soursop, and a decrease of the release of this compound males stop feeding. GC-MS analysis of volatiles identified this compound as α-terpineol. Further olfactometry assays using α-terpineol and soursop, demonstrated that this combination is double attractive to Annonaceae weevils than only soursop volatiles. The results showed a complementation effect between α-terpineol and soursop volatiles. Thus, α-terpineol is the aggregation pheromone of O. palmaris , and its concentration is enhanced by host-plant volatiles.

Reproductive division of labor in insect societies is regulated through multiple concurrent mechanisms, primarily chemical and behavioral. Here, we examined if the Dufour's gland secretion in the primitively eusocial bumble bee Bombus impatiens signals information about caste, social condition, and reproductive status. We chemically analyzed Dufour's gland contents across castes, age groups, social and reproductive conditions, and examined worker behavioral and antennal responses to gland extracts. We found that workers and queens each possess caste-specific compounds in their Dufour's glands. Queens and gynes differed from workers based on the presence of diterpene compounds which were absent in workers, whereas four esters were exclusive to workers. These esters, as well as the total amounts of hydrocarbons in the gland, provided a separation between castes and also between fertile and sterile workers. Olfactometer bioassays demonstrated attraction of workers to Dufour's gland extracts that did not represent a reproductive conflict, while electroantennogram recordings showed higher overall antennal sensitivity in queenless workers. Our results demonstrate that compounds in the Dufour's gland act as caste- and physiology-specific signals and are used by workers to discriminate between workers of different social and reproductive status.

An electronic biosensor for odors was assembled by immobilizing the silk moth Bombyx mori pheromone binding protein (BmorPBP1) on a reduced graphene oxide surface of a field-effect transistor. At physiological pH, the sensor detects the B. mori pheromones, bombykol and bombykal, with good affinity and specificity. Among the other odorants tested, only eugenol elicited a strong signal, while terpenoids and other odorants (linalool, geraniol, isoamyl acetate, and 2-isobutyl-3-methoxypyrazine) produced only very weak responses. Parallel binding assays were performed with the same protein and the same ligands, using the common fluorescence approach adopted for similar proteins. The results are in good agreement with the sensor's responses: bombykol and bombykal, together with eugenol, proved to be strong ligands, while the other compounds showed only poor affinity. When tested at pH 4, the protein failed to bind bombykol both in solution and when immobilized on the sensor. This result further indicates that the BmorPBP1 retains its full activity when immobilized on a surface, including the conformational change observed in acidic conditions. The good agreement between fluorescence assays and sensor responses suggests that ligand-binding assays in solution can be used to screen mutants of a binding protein when selecting the best form to be immobilized on a biosensor.

It is already known that the beagle breed of domestic dogs produces semiochemicals capable of repelling the brown dog tick, Rhipicephalus sanguineus sensu lato (s.l.). With a view to discovering new non-host semiochemicals as tick repellents, we compared the semiochemicals produced by a putative tick-resistant breed of dog, miniature pinscher, with known tick-resistant (beagle) and tick-susceptible (English cocker spaniel) breeds. Two non-host compounds produced by beagles, i.e. 2-hexanone and benzaldehyde, were shown to be present in samples collected from all three breeds. Furthermore, two compounds, 6-methyl-5-hepten-2-one and 1,2,4-trimethylbenzene, were found in higher amounts in samples collected from miniature pinscher dogs. The mean amounts of benzaldehyde, 2-hexanone and 1,2,4-trimethylbenzene were similar for beagles and miniature pinschers (P > 0.05) and higher than the means observed for cocker spaniels (P < 0.05), whereas the mean amount of 6-methyl-5-hepten-2-one produced by miniature pinschers was significantly higher (P < 0.05) than for the other breeds of dogs. In Petri-dish assays with adult R. sanguineus s.l., 6-methyl-5-hepten-2-one was repellent for all observation periods evaluated for the two highest concentrations (0.100 and 0.200 mg.cm -2 , P < 0.01). The obtained results support our hypothesis that miniature pinschers are a tick-resistant dog breed and agree with previous observations of miniature pinschers being the breed least parasitized by ticks. Furthermore, the non-host semiochemical 6-methyl-5-hepten-2-one has potential to be developed for use as a repellent for the protection of susceptible dogs from R. sanguineus s.l. ticks., (Copyright © 2020 Elsevier GmbH. All rights reserved.)

Cyclocephalini beetles of the genus Cyclocephala (Coleoptera: Melolonthidae: Dynastinae) use flowers of some plants as food, shelter, and mating sites. However, little is known about floral scent chemistry involved in this interaction. Here we show that a sesquiterpene alcohol mediates attraction of Cyclocephala paraguayensis Arrow, on bottle gourd flowers, Lagenaria siceraria (Cucurbitaceae). Both males and females started to aggregate on the flowers at twilight; after that, mating began and remained for the entire night. GC-FID/EAD analysis of the L. siceraria floral scent collected in the field revealed that only the major constituent of the airborne volatiles elicited electroantennographic responses on male and female antennae of C. paraguayensis. This compound was identified as (3S,6E)-nerolidol, which was tested in two field trapping trials in Brazil. In the first bioassay, traps baited with nerolidol (mix of isomers) captured significantly more adult C. paraguayensis than control traps. In the second field trial, catches in traps baited with a mixture of isomers or enantiopure nerolidol were significantly higher than captures in control traps, but the treatments did not differ significantly. Analysis from the gut content of adult C. paraguayensis showed the presence of pollen, suggesting that they also use bottle gourd flowers for their nourishment. Taken together, these results suggest that (3S,6E)-nerolidol plays an essential role in the reproductive behavior of C. paraguayensis by eliciting aggregation, mating, and feeding., Competing Interests: The authors declare no conflict of interest.

The strawberry blossom weevil (SBW), Anthonomus rubi, is a major pest in strawberry fields throughout Europe. Traps baited with aggregation pheromone are used for pest monitoring. However, a more effective lure is needed. For a number of pests, it has been shown that the attractiveness of a pheromone can be enhanced by host plant volatiles. The goal of this study was to explore floral volatile blends of different strawberry species (Fragaria x ananassa and Fragaria vesca) to identify compounds that might be used to improve the attractiveness of existing lures for SBW. Floral emissions of F. x a. varieties Sonata, Beltran, Korona, and of F. vesca, were collected by both solid-phase microextraction (SPME) and dynamic headspace sampling on Tenax. Analysis by gas chromatography/mass spectrometry showed the floral volatiles of F. x ananassa. and F. vesca were dominated by aromatic compounds and terpenoids, with 4-methoxybenzaldehyde (p-anisaldehyde) and α-muurolene the major compounds produced by the two species, respectively. Multi-dimensional scaling analyses separated the blends of the two species and explained differences between F. vesca genotypes and, to some degree, variation between F. x ananassa varieties In two-choice behavioral tests, SBW preferred odors of flowering strawberry plants to those of non-flowering plants, but weevils did not discriminate between odors from F. x ananassa and F. vesca flowering plants. Adding blends of six synthetic flower volatiles to non-flowering plants of both species increased the preference of SBW for these over the plants alone. When added individually to non-flowering plants, none of the components increased the preference of SBW, indicating a synergistic effect. However, SBW responded to 1,4-dimethoxybenzene, a major component of volatiles from F. viridis, previously found to synergize the attractiveness of the SBW aggregation pheromone in field studies.

Using gas chromatography (GC) in conjunction with electron impact mass spectrometry and retention-time comparison, 94 compounds, ranging from 2-methyl-2-propenal to octadecanoic acid, were identified in the interdigital secretions of male and female black wildebeests, Connochaetes gnou (also known as the white-tailed gnu). The constituents of these secretions belong to many different compound classes, including hydrocarbons, alcohols, aromatics and aliphatic carbonyl compounds including carboxylic acids as well as carboxylic acid esters. Relatively small quantitative differences were found between the male and female interdigital secretions. It was concluded that these compounds probably do not play a significant role in territorial marking or in chemical communication between males and females of the species, but they could be involved in preserving the remarkably strong attachment between members of social subgroups in black wildebeest populations.

A trapping-based gas liquid microextraction (GLME) method coupled with gas chromatography-mass spectrometry (GC-MS) was utilized to qualitatively and quantitatively characterize semiochemicals in plants. The main GLME extraction efficiency associated parameters (heating temperature and extraction time) were optimized. The results obtained from GLME process were compared with those of steam distillation and ultrasonic extraction, and the recovery, peak number and reproducibility were evaluated by using Thuja koraiensis Nakai as a representative plant. Furthermore, the quantitative performances of the GLME in terms of sample amount, recoveries of spiked standards and correlation were systematically evaluated using standard addition method, which gave a good quantitative ability for all the compounds with squares of correlation coefficient (r 2 ) of higher than 0.99. Finally, the contents of α-pinene, camphene, linalool, α-terpinenol, β-caryophyllene, α-caryophyllene, and totarol in Thuja koraiensis Nakai samples were quantified, and their concentrations (SD, n = 3) were; 0.65 (0.06), 0.62 (0.05), 4.12 (0.15), 0.99 (0.08), 1.11 (0.07), 0.63 (0.04), and 21.91 (0.25) μg g -1 , respectively. It was demonstrated that GLME is a powerful sample preparation technique for quantitative and qualitative analysis of plant semiochemicals., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier B.V. All rights reserved.)

The evolution of pre-zygotic reproductive isolation is a key step in the process of speciation. In many organisms, particularly insects, chemical labels are used as pheromones for species-specific mate recognition. Although an enormous body of knowledge exists regarding the patterns of pheromone chemical ecology, much less is known about the evolutionary processes that underlie the origin of new mating pheromones. Here, we examine case studies that have illuminated the origins of species-specific mating pheromones and suggest future directions for productive research. This article is part of the theme issue 'Signal detection theory in recognition systems: from evolving models to experimental tests'.

House flies (Musca domestica L.) are mechanical vectors of food-borne pathogens including Salmonella spp., Escherichia coli O157:H7, and Shigella spp., resulting in increased risk of diarrheal disease in areas where flies are abundant. Movement of house flies into food crops may be increased by the presence of honeydew-producing insects feeding on these crops. Using gas chromatography-electroantennogram detection (GC-EAD) and gas chromatography-mass spectrometry (GC-MS), volatile odors that elicited house fly antennal response were identified from naval orange (Osbeck) (Sapindales: Rutaceae) and Marsh grapefruit (Macfad.) (Sapindales: Rutaceae) leaves infested with whitefly (Hemiptera: Aleyrodidae) and from whole faba (L.) (Fabales: Fabaceae) bean plants infested with aphids (Hemiptera: Aphididae). Volatiles identified included benzaldehyde, butyl hexanoate, β-caryophyllene, Δ3-carene, (E)-4,8-dimethyl-1,3,7-nonatriene (DMNT), (Z)-3-hexenyl acetate, myrcene, limonene, linalool, and naphthalene. This was followed by semifield bioassays of volatile blends and individual volatiles to determine house fly attraction to these volatiles. Although fly capture rates in the semifield setting were low, benzaldehyde and (Z)-3-hexenyl acetate were consistently attractive to house flies as individual compounds and as components of volatile blends., (© The Author(s) 2019. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

The Mediterranean fruit fly, Ceratitis capitata (Wiedemann) (Diptera: Tephritidae) poses a major threat to fruit and vegetable production in the United States and throughout the world. New attractants and detection methods could improve control strategies for this invasive pest. In this study, we developed a method that combined thin-layer chromatography (TLC) of tea tree essential oil (TTO) ( Melaleuca alternifolia ) with short-range bioassays to isolate attractive kairomones for male C. capitata . After development, the TLC chromatogram indicated that TTO separated into five major spots, designated as zones 1 to 5. When the TLC plate was exposed to flies, zones 1 and 3 were strongly attractive to male C. capitata . To confirm activity, the developed TLC plate was cut into five zones which were then tested in short-range bioassays. Again, flies were observed to aggregate around zones 1 and 3, which corresponded with R f values of 0.93 and 0.59. In addition, zones 1 to 5 were separated using preparative-TLC, and olfactory responses to volatile emissions from the five fractions were quantified by electroantennography (EAG). Highest amplitude EAG responses were recorded with fractions 1 and 3, further supporting the bioactivity of these samples. In conclusion, a TLC-based bioassay system can provide an effective, rapid screening protocol for initial isolation of insect kairomones from complex mixtures such as essential oils or plant extracts. Further analysis of TTO fractions 1 and 3 is needed to identify the specific constituents attractive to male C. capitata .

Elevational gradients affect the production of plant secondary metabolites through changes in both biotic and abiotic conditions. Previous studies have suggested both elevational increases and decreases in host-plant chemical defences. We analysed the correlation of alkaloids and polyphenols with elevation in a community of nine Ficus species along a continuously forested elevational gradient in Papua New Guinea. We sampled 204 insect species feeding on the leaves of these hosts and correlated their community structure to the focal compounds. Additionally, we explored species richness of folivorous mammals along the gradient. When we accounted for Ficus species identity, we found a general elevational increase in flavonoids and alkaloids. Elevational trends in non-flavonol polyphenols were less pronounced or showed non-linear correlations with elevation. Polyphenols responded more strongly to changes in temperature and humidity than alkaloids. The abundance of insect herbivores decreased with elevation, while the species richness of folivorous mammals showed an elevational increase. Insect community structure was affected mainly by alkaloid concentration and diversity. Although our results show an elevational increase in several groups of metabolites, the drivers behind these trends likely differ. Flavonoids may provide figs with protection against abiotic stressors. In contrast, alkaloids affect insect herbivores and may provide protection against mammalian herbivores and pathogens. Concurrent analysis of multiple compound groups alongside ecological data is an important approach for understanding the selective landscape that shapes plant defences.

Piglets are attracted to maternal faeces early in life. Thus, the aim of this study was to identify faecal maternal semiochemicals that attract piglets and evaluate their effects on piglets at weaning. Faecal samples were collected from eight sows during gestation and lactation. Faecal semiochemicals were extracted and identified using solid phase extraction and GC/MS. A total of 26 volatiles were present in lactating and gestating sow faeces. Sows secreted no unique semiochemical after farrowing. However, the concentration of skatole and myristic acid were 2.68 and 1.13 times higher after farrowing. A free-choice preference assessment showed that piglets had a preference for a feeder sprayed with a solution containing skatole and myristic acid. No preference was found when feeders were sprayed with skatole and myristic acid individually. The application of skatole and myristic acid to the feeders of weaned pigs significantly reduced piglet aggression by 30% and tended to increase feeding behaviour by 35% the first 24 h post-weaning. These results suggest that skatole and myristic acid might be acting as a multicomponent maternal signal that attracts piglets and has a calming effect at weaning.

Bed bugs are pests of public health importance due to their relentless biting habits that can lead to allergies, secondary infections and mental health issues. When not feeding on human blood bed bugs aggregate in refuges close to human hosts. This aggregation behaviour could be exploited to lure bed bugs into traps for surveillance, treatment efficacy monitoring and mass trapping efforts, if the responsible cues are identified. The aim of this study was to identify and quantify the bed bug aggregation pheromone. Volatile chemicals were collected from bed bug-exposed papers, which are known to induce aggregation behaviour, by air entrainment. This extract was tested for behavioural and electrophysiological activity using a still-air olfactometer and electroantennography, respectively. Coupled gas chromatography-electroantennography (GC-EAG) was used to screen the extract and the GC-EAG-active chemicals, benzaldehyde, hexanal, (E)-2-octenal, octanal, nonanal, decanal, heptanal, (R,S)-1-octen-3-ol, 3-carene, β-phellandrene, (3E,5E)-octadien-2-one, (E)-2-nonenal, 2-decanone, dodecane, nonanoic acid, 2-(2-butoxyethoxy)ethyl acetate, (E)-2-undecanal and (S)-germacrene D, were identified by GC-mass spectrometry and quantified by GC. Synthetic blends, comprising 6, 16, and 18 compounds, at natural ratios, were then tested in the still-air olfactometer to determine behavioural activity. These aggregation chemicals can be manufactured into a lure that could be used to improve bed bug management.

The composition of defensive secretion produced by metathoracic scent glands was analysed in males and females of the milkweed bug Lygaeus equestris (Heteroptera) using gas chromatography with mass spectrometric detection (GC-MS). The bugs were raised either on cardenolide-containing Adonis vernalis or on control sunflower seeds in order to determine whether the possibility to sequester cardenolides from their host plants would affect the composition of defensive scent-gland secretion. Profiles of the composition of defensive secretions of males and females raised on sunflower were closely similar, with predominant presence of (E)-2-octenal, (E)-2-octen-1-ol, decanal and 3-octen-1-ol acetate. The secretion of bugs raised on A. vernalis was more sexually dimorphic, and some chemicals e.g. (E,E)-2,4-hexadienyl acetate and 2-phenylethyl acetate were dominant in males, but absent in females. Compared to bugs from sunflower, the scent-gland secretion of bugs raised on A. vernalis was characterized by lower overall intensity of the peaks obtained for detected chemicals and by absence of some chemicals that have supposedly antipredatory function ((E)-2-hexenal, (E)-4-oxo-hex-2-enal, 2,4-octadienal). The results suggest that there might be a trade-off between the sequestration of defensive chemicals from host plants and their synthesis in metathoracic scent-glands.

Sexually deceptive orchids typically depend on specific insect species for pollination, which are lured by sex pheromone mimicry. European Ophrys orchids often exploit specific species of wasps or bees with carboxylic acid derivatives. Here, we identify the specific semiochemicals present in O. insectifera , and in females of one of its pollinator species, Argogorytes fargeii . Headspace volatile samples and solvent extracts were analysed by GC-MS and semiochemicals were structurally elucidated by microderivatisation experiments and synthesis. ( Z )-8-Heptadecene and n -pentadecane were confirmed as present in both O. insectifera and A. fargeii female extracts, with both compounds being found to be electrophysiologically active to pollinators. The identified semiochemicals were compared with previously identified Ophrys pollinator attractants, such as ( Z )-9 and ( Z )-12-C 27 -C 29 alkenes in O. sphegodes and ( Z )-9-octadecenal, octadecanal, ethyl linoleate and ethyl oleate in O. speculum , to provide further insights into the biosynthesis of semiochemicals in this genus. We propose that all these currently identified Ophrys semiochemicals can be formed biosynthetically from the same activated carboxylic acid precursors, after a sequence of elongation and decarbonylation reactions in O. sphegodes and O. speculum , while in O. insectifera , possibly by decarbonylation without preceding elongation.

Trail pheromones deposited by ants lead nestmates to food sources. Based on previous evidence that the trail pheromone of the carpenter ant Camponotus modoc originates from the hindgut, our objective in this study was to identify the key component(s) of the pheromone. We collected C. modoc colonies from conifer forests and maintained them in an outdoor enclosure near our laboratory for chemical analyses and behavioral experiments. In gas chromatographic-electroantennographic detection and gas chromatography-mass spectrometric analyses of worker ant hindgut extracts, we identified five candidate components: 2,4-dimethylhexanoic acid, 2,4-dimethyl-5-hexanolide, pentadecane, dodecanoic acid and 3,4-dihydro-8-hydroxy-3,5,7-trimethylisocoumarin. In a series of trail-following experiments, ants followed trails of synthetic 2,4-dimethyl-5-hexanolide, a blend of the five compounds, and hindgut extract over similar distances, indicating that the hexanolide accounted for the entire behavioral activity of the hindgut extract. The hexanolide not only mediated orientation of C. modoc foragers on trails, it also attracted them over distance, indicating a dual function. Further analyses and bioassays with racemic and stereoselectively synthesized hexanolides revealed that the ants produce, and respond to, the (2S,4R,5S)-stereoisomer. The same stereoisomer is a trail pheromone component in several Camponotus congeners, indicating significant overlap in their respective trail pheromone communication systems.

Among major cereals domesticated as staple food, only sorghum has a high proportion of cultivars with condensed tannins in grain, which can trigger bitter taste perception in animals by binding to type 2 taste receptors (TAS2Rs). Here, we report the completion of uncovering of a pair of duplicate recessive genes (Tannin1 and Tannin2) underlying tannin presence. Three loss-of-function alleles from each gene were identified in non-tannin sorghum desired as palatable food. Condensed tannins effectively prevented sparrows from consuming sorghum grain. Parallel geographic distributions between tannin sorghum and Quelea quelea supported the role of tannins in fighting against this major herbivore threat. Association between geographic distributions of human TAS2R variants and tannin sorghum across Africa suggested that different causes had probably driven this bidirectional selection according to varied local herbivore threats and human taste sensitivity. Our investigation uncovered coevolution among humans, plants and environments linked by allelochemicals.

Background: Most allelochemicals are secondary products released from root excretions or plant residues that accumulate in continuous cropping systems and cause severe decline in peanut yield. Resveratrol is a plant-derived stilbene that is released from peanut residues and accumulates in the soil; however, its allelopathic effects on peanut production are overlooked. Effective management solutions need to be developed to relieve allelopathy caused by soil resveratrol. Here, the biodegradation of resveratrol by the fungal endophyte Phomopsis liquidambari was investigated in a mineral salt medium and a soil trial. Resveratrol and its metabolites (produced by degradation by P. liquidambari) were detected by high-performance liquid chromatography-mass spectrometry (HPLC-MS)., Results: Resveratrol released from peanut residues reached a maximum concentration of 0.18 μg g -1 soil in litterbag experiments. Exogenous resveratrol inhibited peanut growth, nodule formation, and soil dehydrogenase activity, and reduced the soil microbial biomass carbon content and bacterial abundance, indicating an allelopathic role in peanut growth. More than 97% of the resveratrol was degraded within 72 and 168 h by P. liquidambari in pure culture and soil conditions, respectively. Resveratrol was first cleaved to 3,5-dihydroxybenzaldehyde and 4-hydroxybenzaldehyde, which were subsequently oxidized into 3,5-dihydroxybenzoic acid and 4-hydroxybenzoic acid, respectively. Fungal resveratrol cleavage oxygenase and the related gene expression were enhanced when P. liquidambari was induced by the resveratrol during the incubation., Conclusion: Our results indicate that the practical application of the fungal endophyte P. liquidambari has strong potential for biodegrading soil resveratrol, which can cause allelopathy in peanut continuous cropping systems. © 2019 Society of Chemical Industry., (© 2019 Society of Chemical Industry.)

Monitoring and collection of fly taxa (Diptera: Calliphoridae; Muscidae; Sarcophagidae) of medical, veterinary, and agricultural importance is often routine practice, providing data on target species presence, distribution and abundance. Collection practices currently involve baited trapping and while an inherent bias accompanying the choice of bait is acknowledged, there is little consistency in bait choice between studies and insufficient assessment of trapping success rates for bait types in current use. This study aimed to examine the effect of bait choice on trapping results for six commonly used bait types; a commercial bait (Envirosafe Fly Attractant, Envirosafe Products) and a combination of mixtures of liver, horse manure and 5% sodium sulfide (Na2S). Trapping success was compared under different seasonal conditions (Summer, Autumn, and Spring) to determine the most attractive bait for calliphorid species, with a secondary comparison of kangaroo versus ox liver occurring under summer conditions. Baits containing Na2S were the most successful in captures of all target taxa, with the addition of manure desirable, yielding increased diversity of taxa. Kangaroo liver baits attracted high numbers of Chrysomya spp. (Robineau-Desvoidy, Diptera: Calliphoridae), while Lucilia spp. (Robineau-Desvoidy, Diptera: Calliphoridae) were comparatively underrepresented in traps using liver alone. The use of a combined ox liver/manure/Na2S bait is recommended as the gold standard for generic screening of necrophagous flies. Where more specific target fly taxa are desired, such as Chrysomya spp. or Lucilia spp. only, alternate baits such as kangaroo or lamb may prove more attractive/successful., (© The Author(s) 2019. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

The present study describes the chemical composition of the essential oil of different plant parts of Devrra tortuosa ; in vivo and in vitro biological activities of plant extract and essential oils. Apiol was found to be the major component of the oil (between 65.73% and 74.41%). The best antioxidant activities were observed for the oil of flowers (IC50 = 175 μg/ml). The samples of stems and roots exhibit lower antioxidant activity (IC50 = 201 μg/ml and 182 μg/ml, respectively). The values of IC50 showed that the extracts of methanol exhibit the highest antioxidants activities (IC50 = 64.8 102 μg/ml). EOs showed excellent antifungal activity against yeasts with low azole susceptibilities (i.e. Malassezia spp. and Candida krusei ). The MIC values of oils varied between 2.85 mg/mL and 27 mg/mL. The obtained results also showed that the plant extracts inhibited the germination and the shoot and root growth of Triticum æstivum seedlings.

The box tree moth, Cydalima perspectalis, is an invasive pest in Europe causing damage on Buxus species. In this study, we aimed to develop a "bisexual" lure to attract both female and male moths. Based on a previous screening bioassay we tested methyl salicylate, phenylacetaldehyde and eugenol as potential attractants in different combinations. The trapping results showed that both binary and ternary blends attracted male and female moths. Catches with these blends were comparable to catches with the synthetic pheromone. Subsequently we carried out single sensillum recordings, which proved the peripheral detection of the above-mentioned compounds on male and female antennae. To identify synergistic flower volatiles, which can be also attractive and can increase the trap capture, we collected flower headspace volatiles from 12 different flowering plant species. Several components of the floral scents evoked good responses from antennae of both females and males in gas chromatography-electroantennographic detection. The most active components were tentatively identified by gas chromatography coupled mass spectrometry as benzaldehyde, cis-ß-ocimene, (±)-linalool and phenethyl alcohol. These selected compounds in combination did not increase significantly the trap capture compared to the methyl salicylate- phenyacetaldehyde blend. Based on these results we discovered the first attractive blend, which was able to attract both adult male and female C. perspectalis in field conditions. These results will yield a good basis for the optimization and development of a practically usable bisexual lure against this invasive pest.

Honey bee (Apis mellifera) queens produce pheromones responsible for mediating both male mating behavior and many critical facets of worker social organization within their colony. These pheromones are dynamic multi-component blends, allowing the communication of detailed information. Indeed, variation in the queen's mating and reproductive state is associated with significant changes in her pheromone profiles, and these different pheromone profiles elicit different behavioral and physiological responses in female workers. Here we evaluate behavioral responses of male drones to the chemical blends produced by two exocrine glands in queens, and determine if the blends and responses are altered by the queen's mating and reproductive state. We find that drone attraction to the chemical blends of mandibular glands produced by mated, laying queens versus virgin queens is reduced, suggesting that the queens produce a reliable signal of their mating receptivity. Interestingly, while the chemical blends of mating, laying queens and virgins queens largely overlap, mated, laying queens produce a greater number of chemicals and greater quantities of certain chemicals than virgin queens, suggesting that these chemicals may serve to inhibit behavioral responses of drones to mated, laying queens. Thus, our results highlight the importance of considering chemical cues and signals that serve to both stimulate and inhibit behavioral responses during social interactions in animals.

Seasonally reproducing animals show many behavioral and physiological changes during the mating period, including increased signaling for intrasexual competition and mate attraction. We collected 102 anogenital gland secretions (AGS) from marking trees in Foping Nature Reserve, and used gas chromatography mass spectrometry analyze these chemical composition. Of these marks, all but one were from males, confirmed with DNA analysis. We found that several chemical constituents, especially volatile compounds, is present only during the mating season and that the relative abundance of many compounds changed as a function of breeding season, whereas nonvolatile compounds were lower in the mating season. This seasonal variation in chemical composition of AGS most likely plays an important role in governing giant panda reproduction, including mate location, attraction, and male-male competition. The chemical properties of many of these putative chemosignals-such as volatility and longevity-are suggestive of these roles, and undoubtedly contribute to successful reproduction for this species with a characteristically sophisticated chemical communication system. We also found a number of important differences between the chemical constituents of AGS from wild pandas and those found in previous studies with captive pandas, suggesting that inappropriate chemosignal composition may contribute to poor reproductive success in captive breeding programs.

Large social insect colonies exhibit a remarkable ability for recognizing group members via colony-specific cuticular pheromonal signatures. Previous work suggested that in some ant species, colony-specific pheromonal profiles are generated through a mechanism involving the transfer and homogenization of cuticular hydrocarbons (CHCs) across members of the colony. However, how colony-specific chemical profiles are generated in other social insect clades remains mostly unknown. Here we show that in the honey bee ( Apis mellifera ), the colony-specific CHC profile completes its maturation in foragers via a sequence of stereotypic age-dependent quantitative and qualitative chemical transitions, which are driven by environmentally-sensitive intrinsic biosynthetic pathways. Therefore, the CHC profiles of individual honey bees are not likely produced through homogenization and transfer mechanisms, but instead mature in association with age-dependent division of labor. Furthermore, non-nestmate rejection behaviors seem to be contextually restricted to behavioral interactions between entering foragers and guards at the hive entrance., Competing Interests: CV, JK, KM, AH, JL, YB No competing interests declared, (© 2019, Vernier et al.)

Abelia chinensis R. Br. (Dipsacales: Caprifoliaceae) is one of the preferred nectar host plants for Culex pipiens pallens Coquillett (Diptera: Culicidae). However, the volatile compounds of its flowers that might be involved in directing mosquitoes' orientation to its nectaries remain unknown. In the present study, the volatile compounds released by A. chinensis florets were collected by solid phase microextraction fiber and analyzed by gas chromatography-mass spectrometry system. Based on the major component species in the volatile profile, a synthetic phytochemical blend (Blend B, composed of six compounds at their most attractive concentrations) was formulated, and its attractiveness was tested against the pentane extract of A. chinensis florets at most attractive concentration (Blend A) and a formerly developed synthetic phytochemical blend (Blend C) in the olfactometer, respectively. The results revealed that the volatile profile of A. chinensis florets was mainly composed of aromatic compounds, most of which had been reported to be attractive to other mosquito species. The synthetic Blend B was as attractive as Blend A (10-1-fold of the crude pentane extract) in the olfactometer bioassays, but they were not as attractive as the formerly developed Blend C. The present study indicated that quantitative and qualitative differences in the constituents of phytochemical blends could significantly affect their attractiveness to Cx. pipiens pallens, and the capture efficiency of phytochemical attractants deserves further research before being applied in the field.

Cyperus alternifolius (C. alternifolius) and Canna generalis (C. generalis) are widely used as artificial floating-bed (AFB) plants for water pollution control. This study evaluated the release of anti-cyanobacterial allelochemicals from both plants in AFB systems. A series of cyanobacterial assays using pure culture solutions and extracts of culture solutions of C. alternifolius and C. generalis demonstrated allelopathic growth inhibition of a cyanobacterium M. aeruginosa. After 45 days of incubation by the culture solutions, both final inhibitory rates of M. aeruginosa were more than 99.6% compared with that of the control groups. GC/MS analyses indicated the presence of a total of 15 kinds of compounds, including fatty acids and phenolic compounds, in both plants' culture solutions, which are are anti-cyanobacterial. These findings provide a basis to apply artificial floating-bed plants for cyanobacterial inhibition using allelopathic effects.

A rapid microwave-assisted extraction and derivatization of three branched chain fatty acids (BCFA): 4-methyloctanoic acid (MOA), 4-ethyloctanoic acid (EOA) and 4-methylnonanoic acid (MNA) from subcutaneous adipose tissue (SAT) of lamb was developed and validated. Linearity was excellent (R 2 > 0.99), and the limits of detection and quantification (μg/mL) were between 0.03 to 0.05 and 0.04 to 0.06, respectively. The relative response factors were 0.9416, 1.2840, and 1.0370 determined with high precision (RSD: 5.8%, 6.7%, and 5.9%) for MOA, EOA, and MNA, respectively. The accuracy, recoveries and matrix effect were 92.5% to 108.3%, 103.1% to 103.8%, and 89.3% to 101.9%, respectively. Analysis of 24 SAT samples can be done in under 4 hr. The levels of total BCFA in SAT of lambs were in good agreement with levels found in literature using more time-consuming method. PRACTICAL APPLICATION: Microwave-assisted preparation and quantification of branched chain fatty acids (BCFA) in lamb subcutaneous adipose tissue (SAT) was developed and validated. This method requires the use of less toxic chemicals and a relatively higher number of samples can be analyzed in a short period of time in comparison to established methods of fatty acid analysis. The validated method met the internationally accepted standards, and the BCFA levels in lamb SAT were consistent with published values; consequently, the method can be used for future analyses., (© 2018 Institute of Food Technologists®.)

Chemical cues are thought to play an important role in mate identification in the solitary giant panda (Ailuropoda melanoleuca). The goal of this study was to detect and identify volatile compounds present in the enclosure air of captive giant pandas. We hypothesized that a subset of compounds produced from breeding animals would be detected in environmental samples because highly volatile chemicals are likely to facilitate mate detection. Samples were collected from the enclosures of 8 giant pandas (n = 4 male, n = 4 female) during the Mar-June breeding season and the Aug-Jan non-breeding period from 2012-2015. Volatile compounds were captured by securing a solid phase micro extraction fiber approximately 3 meters above the ground within a panda enclosure for 6-12 hours. Compounds adsorbed onto the SPME fibers were analyzed by gas chromatography mass spectrometry. Thirty-three compounds were detected in at least 10% of all samples within individual and season and across all subjects within each season. Aromatic compounds made up 27.3% of the enclosure volatile profile, while 21.2% was made of cyclic aliphatic compounds and 51.5% of the enclosure profile was comprised of acyclic aliphatic compounds. Three compounds were likely to be present in male enclosures regardless of season, while Undecane, 4-methyl had a significant (p<0.05) predicted probability of being present in female enclosures. 3,3'-(1,1-Ethanediyl)bis(1H-indole) had a significant (p<0.05) probability of occurrence in male enclosures during the breeding season. Given the prevalence of these compounds, we suspect that these chemicals are important in giant panda communication. This novel sampling technique can detect volatile compounds produced by captive species and also may be a useful tool for detecting pheromones in free-ranging individuals., Competing Interests: The authors have declared that no competing interests exist.

In the Central American region, the aggressive, sibling bark beetles Dendroctonus frontalis Zimmermann (Coleoptera: Curculionidae: Scolytinae) and Dendroctonus mesoamericanus Armendáriz-Toledano & Sullivan (Coleoptera: Curculionidae: Scolytinae) commonly colonize pines concurrently, and in nature they avoid heterospecific pairing, although it can be produced in the lab. We performed walking arrestment bioassays in the lab to examine the capacity of both sexes of both species to discriminate odors from frass expelled from gallery entrances of either solitary females or conspecific pairs of either species. Males of both species strongly preferred odors of frass from solitary, conspecific females over those of heterospecific females or pairs of either species. Female D. frontalis did not discriminate among these frass categories, whereas female D. mesoamericanus preferred frass of conspecific females. In gas chromatography-electroantennographic detection and gas chromatography-mass spectrometry analyses, we determined that males of both species could sense a nearly identical spectrum of approximately 16 host- and beetle-produced compounds present in frass of females of one or both species. Only two of these compounds, endo-brevicomin and ipsdienol, which were present in frass of female D. mesoamericanus and pairs of either species but absent in frass of solitary D. frontalis females, qualitatively distinguished these categories. Several known attractants and synergists for either species declined in concentration postpairing. Our results complement earlier research and indicate how semiochemical composition and concentration in frass might mediate male discrimination of attack sites of conspecific, unpaired females. Furthermore, our data indicate that semiochemical responses for walking females differ from those of males and between species.

Previously, we showed that the symbiotic yeast Lachancea thermotolerans (Filippov) (Saccharomycetales: Saccharomycetaceae) is attractive to its Vespula (Hymenoptera: Vespidae) yellowjacket hosts when grown on media supplemented with grape juice. We hypothesized that "Concerto", a commercial strain of this yeast, could be combined with fruit powder to form a shelf-stable bait for trapping yellowjackets. Using molecular techniques, we first confirmed that Concerto yeast is indeed the species L. thermotolerans. We then tested whether: 1) Concerto yeast produces volatiles similar to those produced by L. thermotolerans isolated from yellowjackets, 2) Concerto yeast enhances attraction of yellowjackets to fruit powder, 3) a Concerto yeast/fruit powder bait interacts synergistically with a yellowjacket semiochemical lure, and 4) a synthetic analog blend of Concerto-produced volatiles attracts yellowjackets. Using gas chromatography-mass spectrometry, we demonstrated that the chemical composition of Concerto-produced volatiles closely resembles that produced by a yellowjacket-isolated strain of L. thermotolerans. In field experiments, addition of Concerto to fruit powder doubled its attractiveness to yellowjackets. Addition of the Concerto/fruit powder bait to a heptyl butyrate-based wasp lure revealed a weak additive effect. A three-component synthetic analog blend of volatiles identified from the Concerto/fruit powder bait attracted Vespula pensylvanica (Saussure), but no other yellowjacket species. Our results suggest that commercial L. thermotolerans in combination with fruit powder could be used as a yellowjacket bait, and that addition of yeast-produced volatiles to a commercial wasp lure may improve its attractiveness to V. pensylvanica. Further research should determine why the synthetic volatile blend failed to attract Vespula species other than V. pensylvanica.

Harmful microalgal blooms are a threat to aquatic organisms, ecosystems and human health. Toxic dinoflagellates of the genus Alexandrium are known to produce paralytic shellfish toxins and to release bioactive extracellular compounds (BECs) with potent cytotoxic, hemolytic, ichtyotoxic and allelopathic activity. Negative allelochemical interactions refer to the chemicals that are released by the genus Alexandrium and that induce adverse effects on the physiology of co-occurring protists and predators. Releasing BECs gives the donor a competitive advantage that may help to form dense toxic blooms of phytoplankton. However BECs released by Alexandrium minutum are uncharacterized and it is impossible to quantify them using classical chemical methods. Allelochemical interactions are usually quantified through population growth inhibition or lytic-activity based bioassays using a secondary target organism. However these bioassays require time (for growth or microalgal counts) and/or are based on lethal effects. The use of pulse amplitude modulation (PAM) fluorometry has been widely used to assess the impact of environmental stressors on phytoplankton but rarely for allelochemical interactions. Here we evaluated the use of PAM and propose a rapid chlorophyll fluorescence based bioassay to quantify allelochemical BECs released from Alexandrium minutum. We used the ubiquitous diatom Chaetoceros muelleri as a target species. The bioassay, based on sub-lethal effects, quantifies allelochemical activity from different samples (filtrates, extracts in seawater) within a short period of time (2 h). This rapid bioassay will help investigate the role of allelochemical interactions in Alexandrium bloom establishment. It will also further our understanding of the potential relationship between allelochemical activities and other cytotoxic activities from BECs. While this bioassay was developed for the species A. minutum, it may be applicable to other species producing allelochemicals and may provide further insights into the role and impact of allelochemical interactions in forming dense algal blooms and structuring marine ecosystems., (Copyright © 2018 Elsevier Ltd. All rights reserved.)