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Foraging ticks reportedly exploit diverse cues to locate their hosts. Here, we tested the hypothesis that host-seeking Western black-legged ticks, Ixodes pacificus , and black-legged ticks, I. scapularis , respond to microbes dwelling in sebaceous gland secretions of white-tailed deer, Odocoileus virginianus , the ticks' preferred host. Using sterile wet cotton swabs, microbes were collected from the pelage of a sedated deer near forehead, preorbital, tarsal, metatarsal and interdigital glands. Swabs were plated on agar, and isolated microbes were identified by 16S rRNA amplicon sequencing. Of 31 microbial isolates tested in still-air olfactometers, 10 microbes induced positive arrestment responses by ticks, whereas 10 others were deterrent. Of the 10 microbes prompting arrestment by ticks, four microbes—including Bacillus aryabhattai (isolates A4)—also attracted ticks in moving-air Y-tube olfactometers. All four of these microbes emitted carbon dioxide and ammonia as well as volatile blends with overlapping blend constituents. The headspace volatile extract (HVE) of B. aryabhattai (HVE-A4) synergistically enhanced the attraction of I . pacificus to CO 2 . A synthetic blend of HVE-A4 headspace volatiles in combination with CO 2 synergistically attracted more ticks than CO 2 alone. Future research should aim to develop a least complex host volatile blend that is attractive to diverse tick taxa.

Background Harnessing insect ecology for insect control is an innovative concept that seeks to exploit, among others, insect-microbe ecological interactions for improved control of pest insects. Microbe-produced cheese odour attracts several dipterans, including host-seeking mosquitoes, but this phenomenon has not been thoroughly explored for mosquito control. Here we tested the hypothesis that attraction of mosquitoes to cheese odour can be exploited as an ecological trap for mosquito control. Results In laboratory and/or field experiments, we show that (i) each of five cheese varieties tested (Raclette, Pecorino, Brie, Gruyere, Limburger) strongly attracts female A. aegypti and Culex. pipiens; (ii) cheese infusions, or headspace odourant extracts (HOEs) of cheese infusions, significantly affect oviposition choices by mosquitoes, (iii) HOEs contain at least 13 odourants; (iv) in field settings, cheese infusions more effectively stimulate mosquito oviposition than positive bluegrass infusion controls, and also capture (by drowning) the spotted wing Drosophila, Drosophila suzukii; and (v) home-made cheese infusions modulate oviposition choices by mosquito females and affect the survivorship of their offspring larvae. Conclusion Our data show that microbial metabolites associated with cheese are attractive to mosquito females seeking hosts and oviposition sites and are likely toxic to mosquito larvae. These microbes and their metabolites could thus be co-opted for both the attract, and the kill, function of 'attract & kill' mosquito control tactics. Implementation of customizable and non-conventional nutritional media as microbe-based ecological traps presents a promising concept which exploits insect ecology for insect control. This article is protected by copyright. All rights reserved.

Four species of Limonius wireworms (Coleoptera: Elateridae), L. californicus, L. canus, L. infuscatus and L. agonus, are serious crop pests in North America. Limoniic acid, (E)-4-ethyloct-4-enoic acid, has been reported as a sex pheromone component of female L. californicus and L. canus, and a sex attractant for male L. infuscatus. In the same study, both limoniic acid and the analog (E)-5-ethyloct-4-enoic acid were highly attractive in field experiments. Moreover, six carboxylic acids in headspace volatiles of Limonius females elicited responses from male antennae but were not tested for behavioral activity. Here, we report trap catch data of Limonius spp. obtained in field experiments at 27 sites across North America. All four Limonius species were attracted to limoniic acid and to the analog but not to the carboxylic acids. Adding these carboxylic acids to limoniic acid, or to the analog, reduced its attractiveness. In dose–response studies, trap lures containing 0.4 mg or 4 mg of limoniic acid afforded large captures of L. californicus and L. infuscatus. Neither limoniic acid nor the analog were deterrent to other elaterid pest species. The broad attractiveness of limoniic acid to Limonius spp., and its non-deterrent effect on heterogeners, may facilitate the development of generic pheromone-based monitoring and management tools for multiple click beetle species.

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.

An underlying assumption of optimal foraging models is that animals are behaviorally, morphologically, and physiologically adapted to maximize their net energy intake. Here we explored whether this concept applies to web-building spiders in a multi-trophic context. If a spider were to build her web next to herbivore-fed-on plants that signal the herbivores’ enemies for help by emitting herbivore-induced plant volatiles (HIPVs), that spider may maximize web captures in the short term. However, she would also risk predation by generalist predators that “listen” to signaling plants to find both herbivore and spider prey, likely resulting in lower overall reproductive fitness for the spider. We tested the hypothesis that HIPVs trigger avoidance responses by web-building spiders. We selected seven common HIPVs and one HIPV elicitor, and in two-choice olfactometer bioassays tested their effect on four synanthropic spider species (false black widow, Steatoda grossa; common cellar spider, Pholcus phalangioides; hobo spider, Eratigena agrestis; western black widow, Latrodectus hesperus). The 8-component HIPV/HIPV elicitor blend had a weak deterrent effect on S. grossa, but the effect did not extend to P. phalangioides, E. agrestis, and L. hesperus. Our findings imply that there was insufficient selection pressure for these spiders to recognize HIPVs in a multi-trophic context, where spiders themselves could become prey if generalist predators or spider-hunting parasitoid wasps were to respond to signaling plants.

Trypodendron retusum (LeConte) (Coleoptera: Curculionidae: Scolytinae) females excised from newly attacked trembling aspen, Populus tremuloides Michaux (Salicaceae), were shown for the first time to produce the aggregation pheromone (+)-lineatin. Coupled gas chromatographic-electroantennographic detection analysis (GC-EAD) disclosed that the antennae of T. retusum, as well as the antennae of three sympatric species, native T. lineatum (Olivier) and T. rufitarsus (Kirby) and exotic T. domesticum (L.), respond to synthetic (+)-lineatin, but not the (−) enantiomer. In contrast, the antennae of T. betulae Swaine responded to SR- and RR-linalool oxide pyranoid and did not detect lineatin. GC-EAD analysis of volatiles from host and nonhost tree species revealed that conifer-produced α-pinene and angiosperm-produced conophthorin and salicylaldehyde were perceived by the antennae of all three native lineatin-perceiving species, suggesting behavioral activity. Field trapping experiments showed that salicylaldehyde synergized the response of coastal, but not interior, T. retusum to lineatin and inhibited the response of T. lineatum and T. rufitarsus. In the absence of salicylaldehyde, α-pinene appeared to inhibit the response of interior T. retusum to lineatin, while for T. lineatum and T. rufitarsus it had an apparent positive additive or synergistic effect. No behavioral response occurred to conophthorin. The results provide evidence for semiochemical-based reproductive isolation between T. retusum and T. betulae, and between these two angiosperm-infesting species and the two conifer-infesting species. They do not explain how isolation could be maintained between T. lineatum and T. rufitarsus.

Mammalian pheromones often linger in the environment and thus are particularly susceptible to interceptive eavesdropping, commonly understood as a one-way dyadic interaction, where prey sense and respond to the scent of a predator. Here, we tested the “counterespionage” hypothesis that predator and prey co-opt each other’s pheromone as a cue to locate prey or evade predation. We worked with wild brown rats (predator of mice) and wild house mice (prey of brown rats) as model species, testing their responses to pheromone-baited traps at infested field sites. The treatment trap in each of two trap pairs per replicate received sex attractant pheromone components (including testosterone) of male mice or male rats, whereas corresponding control traps received only testosterone, a pheromone component shared between mouse and rat males. Trap pairs disseminating male rat pheromone components captured 3.05 times fewer mice than trap pairs disseminating male mouse pheromone components, and no female mice were captured in rat pheromone-baited traps, indicating predator aversion. Indiscriminate captures of rats in trap pairs disseminating male rat or male mouse pheromone components, and fewer captures of rats in male mouse pheromone traps than in (testosterone-only) control traps indicate that rats do eavesdrop on the male mouse sex pheromone but do not exploit the information for mouse prey location. The counterespionage hypothesis is supported by trap catch data of both mice and rats but only the mice data are in keeping with our predictions for motive of the counterespionage.

Formicine ants in distress spray alarm pheromone which typically recruits nestmates for help. Studying the western carpenter ant, Camponotus modoc Wheeler (Hymenoptera: Formicidae), our objectives were to (1) determine the exocrine glands that contain alarm recruitment pheromone, (2) identify the key alarm recruitment pheromone components, and (3) ascertain the pheromone components that are discharged by distressed ants. In Y‐tube olfactometer experiments, extracts of poison glands, but not of Dufour’s glands, elicited anemotactic responses from worker ants. Gas chromatographic‐mass spectrometric analyses of poison gland extracts revealed the presence of (1) aliphatic alkanes (undecane, tridecane, pentadecane, heptadecane), (2) aliphatic alkenes [(Z)‐7‐pentadecene, (Z)‐7‐ and (Z)‐8‐heptadecene], (3) two acids (formic, benzoic), and (4) other oxygenated compounds (hexadecan‐1‐ol, hexadecyl formate, hexadecyl acetate). Testing the responses of worker ants in Y‐tube olfactometers to complete and partial synthetic blends of these compounds revealed that the acids and the alkanes are essential alarm pheromone components. In two‐choice arena bioassays, micro‐locations treated with synthetic alarm pheromone recruited worker ants. Acids and alkanes were abundant in the poison gland and the Dufour’s gland, respectively, suggesting that the alarm pheromone components originate from both glands. Moreover, alarm pheromone sprays of ants differed in that all sprays contained formic acid but only some also contained alkanes, implying that ants can independently discharge the content of either one or both glands in accordance with the type of distress incident they experience.

Four species of Tetramorium pavement ants are known to guide foraging activities of nestmates via trail pheromones secreted from the poison gland of worker ants, but the trail pheromone of T. immigrans is unknown. Our objectives were to (1) determine whether poison gland extract of T. immigrans workers induces trail-following behavior of nestmates, (2) identify the trail pheromone, and (3) test whether synthetic trail pheromone induces trail-following behavior of workers. In laboratory no-choice bioassays, ants followed poison-gland-extract trails farther than they followed whole-body-extract trails or solvent-control trails. Gas chromatographic-electroantennographic detection (GC-EAD) analyses of poison gland extract revealed a single candidate pheromone component (CPC) that elicited responses from worker ant antennae. The CPC mass spectrum indicated, and an authentic standard confirmed, that the CPC was methyl 2-methoxy-6-methylbenzoate (MMMB). In further laboratory no-choice bioassays, ants followed poison-gland-extract trails (tested at 1 ant equivalent) and synthetic MMMB trails (tested at 0.35 ant equivalents) equally far, indicating that MMMB is the single-component trail pheromone of T. immigrans. Moreover, in laboratory two-choice bioassays, ants followed MMMB trails ~ 21-times farther than solvent-control trails. In field settings, when T. immigrans colonies were offered a choice between two paper strips treated with a synthetic MMMB trail or a solvent-control trail, each leading to an apple bait, the MMMB trails efficiently recruited nestmates to baits.

As part of an ongoing program to identify sex attractant pheromone components that mediate sexual communication in yellowjacket wasps, a novel sesquiterpene was isolated from body surface extracts of virgin bald-faced hornet queens, Dolichovespula maculata. The gross structure of this sesquiterpene was proposed through microscale spectroscopic analyses, and the configuration of the central olefin was subsequently confirmed by total synthesis. This new natural product (termed here dolichovespulide) represents an important addition to the relatively small number of terpenoids reported from the taxonomic insect family Vespidae.

Plant essential oils (EOs) have been considered as spatial repellents to help disrupt the pathogen transmission cycle of mosquitoes. Our objective was to assess spatial repellency effects of EOs on the tropical yellow fever mosquito, Aedes aegypti (L.) (Diptera: Culicidae) and on local mosquito populations in coastal British Columbia (Canada). In laboratory experiments using protocols of the World Health Organization, three of the solitary EOs tested proved repellent to Ae. aegypti: cinnamon bark, lemongrass, and rosemary. Binary combinations of select EOs enhanced the repellent effect of single EOs through synergistic interactions. The EO blend of geranium and peppermint lowered the RD50 (the dose required to obtain 50% repellency) of each solitary EO by >1,000-fold. Compared with binary EO blends, ternary EO blends were typically less repellent to mosquitoes, possibly due to a dilution effect of the most effective EO constituent(s) in the blend. In field experiments, the EO blend of lemongrass and cinnamon bark expressed spatial repellency towards the cool weather mosquito, Culiseta incidens (Thomson) (Diptera: Culicidae), even when this blend was disseminated from devices as much as 1 m away from a sentinel trap releasing attractive vertebrate host odorants and CO2. Deployment of EOs as spatial repellents in small outdoor gatherings could help protect humans from mosquito-borne diseases, particularly when this tactic is coupled with other tools of mosquito management.

Background There is widespread interkingdom signalling between insects and microbes. For example, microbes found in floral nectar may modify its nutritional composition and produce odorants that alter the floral odor bouquet which may attract insect pollinators. Mosquitoes consume nectar and can pollinate flowers. We identified microbes isolated from nectar of common tansy, Tanacetum vulgare, elucidated the microbial odorants, and tested their ability to attract the common house mosquito, Culex pipiens. Results We collected 19 microbial isolates from T. vulgare nectar, representing at least 12 different taxa which we identified with 16S or 26S rDNA sequencing as well as by biochemical and physiological tests. Three microorganisms (Lachancea thermotolerans, Micrococcus lactis, Micrococcus luteus) were grown on culture medium and tested in bioassays. Only the yeast L. thermotolerans grown on nectar, malt extract agar, or in synthetic nectar broth significantly attracted Cx. pipiens females. The odorant profile produced by L. thermotolerans varied with the nutritional composition of the culture medium. All three microbes grown separately, but presented concurrently, attracted fewer Cx. pipiens females than L. thermotolerans by itself. Conclusions Floral nectar of T. vulgare contains various microbes whose odorants contribute to the odor profile of inflorescences. In addition, L. thermotolerans produced odorants that attract Cx. pipiens females. As the odor profile of L. thermotolerans varied with the composition of the culture medium, we hypothesize that microbe odorants inform nectar-foraging mosquitoes about the availability of certain macro-nutrients which, in turn, affect foraging decisions by mosquitoes.

We tested the hypothesis that the “ecological trap” phenomenon (a mismatch between a habitat’s perceived attractiveness and its actual quality, resulting in a population sink) is exploitable for pest control. We selected mosquitoes as modal organisms, because selection of an oviposition site by adult female mosquitoes in response to its perceived attractiveness is of paramount importance for the development and survival of their larval offspring. In laboratory and/or field experiments, we show that (i) each of five cheese varieties tested (Raclette, Pecorino, Brie, Gruyere, Limburger) strongly attracts females of both the yellow fever mosquito,Aedes aegypti, and the common house mosquito,Cx. pipiens;(ii) cheese infusions, or headspace odorant extracts (HOEs) of cheese infusions, significantly affect oviposition choices byCx. pipiensandAe. aegypti, (iii) HOEs contain at least 13 odorants; (iv) in field settings, cheese infusions more effectively stimulate oviposition byCx. pipiensandCuliseta incidensthan bluegrass (Poasp.) infusions, and also capture (by drowning) the spotted wingDrosophila(SWD); (v) the microbe composition of home-made cheese infusions modulates oviposition choices by mosquitoes; and (vi) the type of cheese infusion coupled with its nutritional content strongly affects the survivorship of mosquito larvae. In combination, our data show that microbial metabolites associated with cheese and cheese infusions are both attractive to adult mosquitoes seeking hosts and oviposition sites, respectively, and are toxic to mosquito larvae. These microbes and their metabolites could thus be coopted for both the attract and the kill function of “attract & kill” mosquito control tactics. Implementation of customizable and non-conventional nutritional media, such as home-made cheese infusions, as microbe-based ecological traps presents a promising concept which exploits insect ecology for insect control.

Wireworms, the larvae of click beetles (Coleoptera: Elateridae), are soil-dwelling insect pests inflicting major economic damage on many types of agricultural crops worldwide. The objective of this work was to identify the female-produced sex pheromones of the Pacific Coast wireworm, Limonius canus LeConte, and the sugarbeet wireworm, L. californicus (Mannerheim) (Coleoptera: Elateridae). Headspace volatiles from separate groups of female L. canus and L. californicus were collected on Porapak Q and analyzed by gas chromatography with electroantennographic detection (GC-EAD) and GC-mass spectrometry. GC-EAD recordings revealed strong responses from male L. canus and male L. californicus antennae to the same compound, which appeared below GC detection threshold. The structure of this candidate pheromone component was deduced from the results of micro-analytical treatments of extracts, retention index calculations on four GC columns, and by syntheses of more than 25 model compounds which were assessed for their GC retention characteristics and electrophysiological activity. The EAD-active compound was identified as (E)-4-ethyloct-4-enoic acid, which we name limoniic acid. In field experiments in British Columbia and Alberta, Canada, traps baited with synthetic limoniic acid captured large numbers of male Limonius click beetles, whereas unbaited control traps captured few. Compared to traps baited with the analogue, (E)-5-ethyloct-4-enoic acid, traps baited with limoniic acid captured 9-times more male L. californicus, and 6.5-times more male western field wireworms, L. infuscatus Motschulsky, but 2.3-times fewer male L. canus. Limoniic acid can now be developed for detection, monitoring and possibly control of L. californicus, L. infuscatus and L. canus populations.

Upon harvest, Western redcedar (WRC, Thuja plicata) trees have a high incidence and extent of heartwood rot. While monoterpenoids and lignans have been linked to rot resistance in this species, other specialized metabolites, such as diterpenes, are likely to contribute to rot resistance. Here we report the cloning and functional assessment of three putative diterpene synthase (TpdiTPS) genes expressed during heartwood formation in WRC. The predicted proteins of the three genes lack either of the two catalytically independent active sites typical of most diTPS, indicating monofunctional rather than bifunctional activity. To identify potential catalytic activities of these proteins, we expressed them in genetically engineered Escherichia coli strains that produce four potential substrates, geranylgeranyl diphosphate (GGDP), ent, syn, and normal stereoisomers of copalyl diphosphate (CDP). We found that TpdiTPS3 used GGDP to produce CDP. TpdiTPS2 used normal CDP to produce levopimaradiene. TpdiTPS1 showed stereoselectivity as it used normal CDP to produce sandaracopimaradiene and syn-CDP to produce syn-stemod-13(17)-ene. These genes and protein enzymatic activities have not been previously reported in WRC and provide an opportunity to assess their potential roles in heartwood rot resistance in this economically important species.

We tested the recent hypothesis that the "fly factor" phenomenon (food currently or previously fed on by flies attracts more flies than the same type of food kept inaccessible to flies) is mediated by bacterial symbionts deposited with feces or regurgitated by feeding flies. We allowed laboratory-reared black blow flies, Phormia regina (Meigen), to feed and defecate on bacterial Luria-Bertani medium solidified with agar, and isolated seven morphologically distinct bacterial colonies. We identified these using matrix-assisted laser desorption/ionization mass spectrometry and sequencing of the 16S rRNA gene. In two-choice laboratory experiments, traps baited with cultures of Proteus mirabilis Hauser, Morganella morganii subsp. sibonii Jensen, or Serratia marcescens Bizio, captured significantly more flies than corresponding control jars baited with tryptic soy agar only. A mixture of seven bacterial strains as a trap bait was more attractive to flies than a single bacterial isolate (M. m. sibonii). In a field experiment, traps baited with agar cultures of P. mirabilis and M. m. sibonii in combination captured significantly more flies than traps baited with either bacterial isolate alone or the agar control. As evident by gas chromatography-mass spectrometry, the odor profiles of bacterial isolates differ, which may explain the additive effect of bacteria to the attractiveness of bacterial trap baits. As "generalist bacteria," P. mirabilis and M. m. sibonii growing on animal protein (beef liver) or plant protein (tofu) are similarly effective in attracting flies. Bacteria-derived airborne semiochemicals appear to mediate foraging by flies and to inform their feeding and oviposition decisions.

Yellowjackets in the genera Vespula and Dolichovespula are prevalent eusocial insects of great ecological and economic significance, but the chemical signals of their sexual communication systems have defied structural elucidation. Herein, we report the identification of sex attractant pheromone components of virgin bald-faced hornet queens (Dolichovespula maculata). We analyzed body surface extracts of queens by coupled gas chromatographic-electroantennographic detection (GC-EAD), isolated the compounds that elicited responses from male antennae by high-performance liquid chromatography (HPLC), and identified these components by GC mass spectrometry (MS), HPLC-MS, and NMR spectroscopy. In laboratory olfactometer experiments, synthetic (2Z,7E)-3,7-dimethyldeca-2,7-diendioic acid (termed here maculatic acid A) and (2Z,7E)-10-methoxy-3,7-dimethyldeca-10-oxo-deca-2,7-dienoic acid (termed here maculatic acid C) in binary combination significantly attracted bald-faced hornet males. These are the first sex attractant pheromone components identified in yellowjackets.

Cuticular hydrocarbons (CHCs) of social insects have typically been studied for their roles in reproductive signaling (i.e., fertility) rather than sexual signaling (i.e., interest in mating), resulting in little information about CHCs of males and virgin females. This dearth of information applies particularly to social wasps. We tested the hypothesis that CHCs differentiate sex, caste, and nest membership in each of four yellowjacket species (baldfaced hornets, Dolichovespula maculata; southern yellowjackets, Vespula squamosa; western yellowjackets, V. pensylvanica; V. alascensis). Cold-euthanized queens (21), gynes (81), workers (125), and males (77) from 35 nests were extracted with pentane, and each of the resulting 304 extracts was analyzed by gas chromatography (GC) and GC–mass spectrometry to identify and quantify CHC constituents (aliphatic alkanes and alkenes; mono-, di-, and tri-methyl-branched alkanes). To determine whether caste and sex differ in CHC profiles of wasps, linear discriminant analyses were performed, using Z-transformed relative CHC peak areas as predictor variables and sex and caste, or nest, as grouping variables. When caste and sex were used as a grouping variable, plots of the first two discriminant functions revealed that wasps from each of the four species clustered into their respective groups (queens, gynes, workers, males), with significant differences in group centroids, as measured by Wilks’ lambda. When nest was used as a grouping variable, plots of the first two discriminant functions revealed that workers from each of the four species and males from each of three species (insufficient sample size for V. pensylvanica) clustered according to nest. Diagnostic power calculations show greater inter-caste than inter-nest variation. Our data support the above hypothesis and inspire future studies to determine the definitive role(s) that gyne- and male-specific CHCs play in the context of sexual communication, from the perspective of both males and females.

Western skunk cabbage, Lysichiton americanus (Araceae), is pollinated mainly by the rove beetle Pelecomalium testaceum (Staphylinidae). Our objective was to determine the floral semiochemical(s) of L. americanus that attract(s) P. testaceum. Porapak Q headspace volatile extracts of L. americanus inflorescences were analyzed by gas chromatographic–electroantennographic detection (GC–EAD) and GC–mass spectrometry. In GC–EAD analyses, three floral odorants [(E)-4 nonene, (E)-5-undecene, indole] elicited consistent responses from the antennae of female P. testaceum. In field experiments, traps baited with a blend of these three components (“3-CB”) captured significantly more P. testaceum than unbaited control traps. Traps baited with the 3-CB, the two hydrocarbons, or indole, each captured significantly more beetles than unbaited control traps, indicating redundancy in the semiochemical blend. Moreover, traps baited with indole captured significantly more beetles than traps baited with either the 3-CB, or the hydrocarbons, indicating that indole is a key floral attractant for P. testaceum. Many necrophilous and coprophilous insects respond to indole in search of carrion or feces, but P. testaceum has never been associated with these types of resources. Both electrophysiological and behavioral responses of P. testaceum to two hydrocarbon semiochemicals, which are not signature odorants of carrion or feces, may indicate that the beetles recognize the odor of L. americanus as an honest signal, seek and pollinate its inflorescences, and get rewarded with pollen and on-plant mating opportunities.

We have recently developed new food baits (SFU Mouse Bait, SFU Rat Bait) for trapping granivorous house mice, Mus musculus, and black rats, Rattus rattus, and for trapping omnivorous brown rats, Rattus norvegicus. Both baits contain synthetic long-range volatile food attractants that represent favourite rodent foods. They draw foraging rodents to the baits where feeding stimulants induce feeding on them. Our objectives were to test the SFU Baits in comparison to three commercial mouse and rat baits, and peanut butter on trap captures of wild house mice, black rats and brown rats. In paired-trap experiments, traps baited with the SFU Mouse Bait captured (i) 6.6 times more mice than Provoke® Mouse (P

Ants select sustained carbohydrate resources, such as aphid honeydew, based on many factors including sugar type, volume and concentration. We tested the hypotheses (H1–H3) that western carpenter ants, Camponotus modoc, seek honeydew excretions from Cinara splendens aphids based solely on the presence of sugar constituents (H1), prefer sugar solutions containing aphid-specific sugars (H2) and preferentially seek sugar solutions with higher sugar content (H3). We further tested the hypothesis (H4) that workers of both Ca. modoc and European fire ants, Myrmica rubra , selectively consume particular mono-, di- and trisaccharides. In choice bioassays with entire ant colonies, sugar constituents in honeydew (but not aphid-specific sugar) as well as sugar concentration affected foraging decisions by Ca. modoc . Both Ca. modoc and M. rubra foragers preferred fructose to other monosaccharides (xylose, glucose) and sucrose to other disaccharides (maltose, melibiose, trehalose). Conversely, when offered a choice between the aphid-specific trisaccharides raffinose and melezitose, Ca. modoc and M. rubra favoured raffinose and melezitose, respectively. Testing the favourite mono-, di- and trisaccharide head-to-head, both ant species favoured sucrose. While both sugar type and sugar concentration are the ultimate cause for consumption by foraging ants, strong recruitment of nest-mates to superior sources is probably the major proximate cause.

Urine of male house mice, Mus musculus, is known to have primer pheromone effects on the reproductive physiology of female mice. Urine-mediated releaser pheromone effects that trigger certain behavioral responses are much less understood, and no field studies have investigated whether urine deposits by male or female mice, or synthetic mouse pheromone, increase trap captures of mice. In field experiments, we baited traps with bedding soiled with urine and feces of caged female or male mice, and recorded captures of mice in these and in control traps containing clean bedding. Traps baited with female bedding preferentially captured adult males, whereas traps baited with male bedding preferentially captured juvenile and adult females, indicating the presence of male- and female-specific sex pheromones in soiled bedding. Analyses of headspace volatiles emanating from soiled bedding by gas chromatography/mass spectrometry revealed that 3,4-dehydro-exo-brevicomin (DEB) was seven times more prevalent in male bedding and that 2-sec-butyl-4,5-dihydrothiazole (DHT) was male-specific. In a follow-up field experiment, traps baited with DEB and DHT captured 4 times more female mice than corresponding control traps, thus indicating that DEB and DHT are sex attractant pheromone components of house mouse males. Our study provides impetus to identify the sex attractant pheromone of female mice, and to develop synthetic mouse pheromone as a lure to enhance the efficacy of trapping programs for mouse control.

Ants deposit trail pheromones that guide nestmates to food sources. We tested the hypotheses that ant community members (Western carpenter ants, Camponotus modoc, black garden ants, Lasius niger, European fire ants, Myrmica rubra) (1) sense, and follow, each other&rsquo, s trail pheromones, and (2) fail to recognize trail pheromones of allopatric ants (pavement ants, Tetramorium caespitum, desert harvester ants, Novomessor albisetosus, Argentine ants, Linepithema humilis). In gas chromatographic-electroantennographic detection analyses of a six-species synthetic trail pheromone blend (6-TPB), La. niger, Ca. modoc, and M. rubra sensed the trail pheromones of all community members and unexpectedly that of T. caespitum. Except for La. niger, all species did not recognize the trail pheromones of N. albisetosus and Li. humilis. In bioassays, La. niger workers followed the 6-TPB trail for longer distances than their own trail pheromone, indicating an additive effect of con- and hetero-specific pheromones on trail-following. Moreover, Ca. modoc workers followed the 6-TPB and their own trail pheromones for similar distances, indicating no adverse effects of heterospecific pheromones on trail-following. Our data show that ant community members eavesdrop on each other&rsquo, s trail pheromones, and that multiple pheromones can be combined in a lure that guides multiple species of pest ants to lethal food baits.

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.

As part of an ongoing program to identify sex attractant pheromone components that mediate sexual communication in yellowjacket wasps, a novel sesquiterpene was isolated from body surface extracts of virgin bald-faced hornet queens

A correction to this article has been published and is linked from the HTML and PDF versions of this paper. The error has been fixed in the paper.

Female mosquitoes exploit olfactory, CO2, visual, and thermal cues to locate vertebrate hosts. Male and female mosquitoes also consume floral nectar that provides essential energy for flight and survival. Heretofore, nectar-foraging mosquitoes were thought to be guided solely by floral odorants. Using common tansies, Tanacetum vulgare L., northern house mosquitoes, Culex pipiens L., and yellow fever mosquitoes, Aedes aegypti (L.), we tested the hypothesis that the entire inflorescence Gestalt of olfactory, CO2 and visual cues is more attractive to mosquitoes than floral odorants alone. In laboratory experiments, we demonstrated that visual and olfactory inflorescence cues in combination attract more mosquitoes than olfactory cues alone. We established that tansies become net producers of CO2 after sunset, and that CO2 enhances the attractiveness of a floral blend comprising 20 synthetic odorants of tansy inflorescences. This blend included nine odorants found in human headspace. The “human-odorant-blend” attracted mosquitoes but was less effective than the entire 20-odorant floral blend. Our data support the hypothesis that the entire inflorescence Gestalt of olfactory, CO2 and visual cues is more attractive to mosquitoes than floral odorants alone. Overlapping cues between plants and vertebrates support the previously postulated concept that haematophagy of mosquitoes may have arisen from phytophagy.

There is widespread interkingdom signalling between insects and microbes. For example, microbes found in floral nectar may modify its nutritional composition and produce odorants that alter the floral odor bouquet which may attract insect pollinators. Mosquitoes consume nectar and can pollinate flowers. We identified microbes isolated from nectar of common tansy, Tanacetum vulgare, elucidated the microbial odorants, and tested their ability to attract the common house mosquito, Culex pipiens.We collected 19 microbial isolates from T. vulgare nectar, representing at least 12 different taxa which we identified with 16S or 26S rDNA sequencing as well as by biochemical and physiological tests. Three microorganisms (Lachancea thermotolerans, Micrococcus lactis, Micrococcus luteus) were grown on culture medium and tested in bioassays. Only the yeast L. thermotolerans grown on nectar, malt extract agar, or in synthetic nectar broth significantly attracted Cx. pipiens females. The odorant profile produced by L. thermotolerans varied with the nutritional composition of the culture medium. All three microbes grown separately, but presented concurrently, attracted fewer Cx. pipiens females than L. thermotolerans by itself.Floral nectar of T. vulgare contains various microbes whose odorants contribute to the odor profile of inflorescences. In addition, L. thermotolerans produced odorants that attract Cx. pipiens females. As the odor profile of L. thermotolerans varied with the composition of the culture medium, we hypothesize that microbe odorants inform nectar-foraging mosquitoes about the availability of certain macro-nutrients which, in turn, affect foraging decisions by mosquitoes.

Trapping brown rats is challenging because they avoid newly placed traps in their habitat. Herein, we report the identification of the sex pheromone produced by male brown rats and its effect on trap captures of wild female brown rats. Collecting urine- and feces-soiled bedding material of laboratory-kept rats and comparing the soiled-bedding odorants of juvenile and adult males, as well as of adult males and females, we found nine compounds that were specific to, or most prevalent in, the odor profiles of sexually mature adult males. When we added a synthetic blend of six of these compounds (2-heptanone, 4-heptanone, 3-ethyl-2-heptanone, 2-octanone, 2-nonanone, 4-nonanone) to one of two paired food-baited trap boxes, these boxes attracted significantly more laboratory-strain female rats in laboratory experiments, and captured ten times more wild female rats in a field experiment than the corresponding control boxes. Our data show that the pheromone facilitates captures of wild female brown rats.

The sex pheromone of the endoparasitoid insect Xenos peckii (Strepsiptera: Xenidae) was recently identified as (7E,11E)-3,5,9,11-tetramethyl-7,11-tridecadienal. Herein we report the asymmetric synthesis of three candidate stereostructures for this pheromone using a synthetic strategy that relies on an sp(3) -sp(2) Suzuki-Miyaura coupling to construct the correctly configured C7-alkene function. Comparison of (1) H NMR spectra derived from the candidate stereostructures to that of the natural sex pheromone indicated a relative configuration of (3R*,5S*,9R*). Chiral gas chromatographic (GC) analyses of these compounds supported an assignment of (3R,5S,9R) for the natural product. Furthermore, in a 16-replicate field experiment, traps baited with the synthetic (3R,5S,9R)-enantiomer alone or in combination with the (3S,5R,9S)-enantiomer captured 23 and 18 X. peckii males, respectively (mean±SE: 1.4±0.33 and 1.1±0.39), whereas traps baited with the synthetic (3S,5R,9S)-enantiomer or a solvent control yielded no captures of males. These strong field trapping data, in combination with spectroscopic and chiral GC data, unambiguously demonstrate that (3R,5S,9R,7E,11E)-3,5,9,11-tetramethyl-7,11-tridecadienal is the X. peckii sex pheromone.

Key message The monoterpenoid content ofThuja plicataneedles and scales differs both quantitatively and qualitatively. Resin storage structures are associated with anatomical modifications that suggest facilitated exit of monoterpenoids.

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.