Behavioral and Chemical Investigations of Contact Kairomones Released by the Mud Dauber Wasp <Emphasis Type="Italic">Trypoxylon politum</Emphasis>, a Host of the Parasitoid <Emphasis Type="Italic">Melittobia digitata</Emphasis>

Contact kairomones from the host mud dauber wasp Trypoxylon politum Say (Hymenoptera: Crabronidae) that mediate behavioral responses of its ectoparasitoid Melittobia digitata Dahms (Hymenoptera: Eulophidae) were investigated. Chemical residues from host by-products, the cocoon, and the meconium, induced arrestment behavior of macropterous female parasitoids, while those from the host stage attacked, i.e., the prepupa, did not. Melittobia digitata response to polar and apolar extracts of host by-products indicated kairomone(s) solubility mainly in hexane. GC and GC/MS analysis of cocoon and meconium apolar extracts revealed a mixture of linear carboxylic acids from C₆ to C₁₈, and both extracts contained almost identical compounds. When a reconstructed blend of host by-product carboxylic acids was tested, M. digitata females showed only a weak response, thus suggesting that other unidentified compounds present in small quantities also may be involved. Melittobia digitata’s response to contact kairomones was innate and not affected by previous host exposure experience. Our results provide evidence of contact kairomone exploitation in the genus Melittobia. The ecological significance of these findings in the host selection process of M. digitata is discussed.     

Cuticular Hydrocarbon Cues Are Used for Host Acceptance by <Emphasis Type="Italic">Pseudacteon</Emphasis> spp. Phorid Flies that Attack <Emphasis Type="Italic">Azteca sericeasur</Emphasis> Ants

Parasitoids often use complex cues to identify suitable hosts in their environment. Phorid fly parasitoids that develop on one or a few host species often use multiple cues, ranging from general to highly specific, to home in on an appropriate host. Here, we describe the hierarchy of cues that Pseudacteon phorid flies use to identify Azteca ant hosts. We show, through behavioral observations in the field, that phorid flies are attracted to two cryptic Azteca species, but only attack Azteca sericeasur (Hymenoptera: Formicidae: Dolichoderinae). To test whether the phorid flies use cuticular hydrocarbons (CHCs) to distinguish between the two Azteca taxa, we first documented and compared cuticular hydrocarbons of the two Azteca taxa using gas chromatography/mass spectrometry. Then, using cuticular hydrocarbon-transfer experiments with live ants, we characterized the cuticular hydrocarbons of A. sericeasur as a short-range, host location cue used by P. lasciniosus (Diptera: Phoridae) to locate the ants.     

<Emphasis Type="Italic">Vitex agnus-castus</Emphasis> is a Preferred Host Plant for <Emphasis Type="Italic">Hyalesthes obsoletus</Emphasis>

Hyalesthes obsoletus Signoret (Homoptera: Cixiidae) is a polyphagous planthopper that transmits stolbur phytoplasma (a causative agent of yellows disease) to various weeds, members of the Solanaceae, and wine grapes (Vitis vinifera L.) in Europe and the Middle East. Planthoppers were collected by hand vacuuming eight native plant species. Vitex agnus-castus L., a shrub in the Verbenaceae, hosted the largest number of H. obsoletus, although Olea europaea L. also served as a host for adults. Using a Y-olfactometer, we compared the planthoppers relative preference for V. agnus-castus, Convolvulus arvensis, and V. vinifera. V. agnus-castus was more attractive to both male and female H. obsoletus than the other plants. H. obsoletus antennal response was stronger to volatiles collected from V. agnus-castus than from Cabernet Sauvignon variety of V. vinifera. To determine if V. agnus-castus would serve as a reservoir for the pathogen, H. obsoletus were collected from leaf and stem samples of native V. agnus-castus, and were tested by polymerase chain reaction (PCR) for the presence of phytoplasma DNA. While 14% and 25% (2003 and 2004, respectively) of the insects tested positive for phytoplasma DNA, none of the plant samples tested positive. To determine if V. agnus-castus could serve as a host plant for the development of the planthopper, we placed emergence cages beneath field shrubs and enclosed wild-caught H. obsoletus in a cage with a potted young shrub. We found adult H. obsoletus in the emergence cases and planthopper nymphs in the soil of the potted plant. We concluded that V. agnus-castus is attractive to H. obsoletus, which seems to be refractory to phytoplasma infections and warrants further testing as a trap plant near vineyards.     

Yellow-Cedar, <Emphasis Type="Italic">Callitropsis</Emphasis> (<Emphasis Type="Italic">Chamaecyparis</Emphasis>) <Emphasis Type="Italic">nootkatensis</Emphasis>, Secondary Metabolites,Biological Activities,and Chemical Ecology

Yellow-cedar, Callitropsis nootkatensis, is prevalent in coastal forests of southeast Alaska, western Canada, and inland forests along the Cascades to northern California, USA. These trees have few microbial or animal pests, attributable in part to the distinct groups of biologically active secondary metabolites their tissues store for chemical defense. Here we summarize the new yellow-cedar compounds identified and their biological activities, plus new or expanded activities for tissues, extracts, essential oils and previously known compounds since the last review more than 40 years ago. Monoterpene hydrocarbons are the most abundant compounds in foliage, while heartwood contains substantial quantities of oxygenated monoterpenes and oxygenated sesquiterpenes, with one or more tropolones. Diterpenes occur in foliage and bark, whereas condensed tannins have been isolated from inner bark. Biological activities expressed by one or more compounds in these groups include fungicide, bactericide, sporicide, acaricide, insecticide, general cytotoxicity, antioxidant and human anticancer. The diversity of organisms impacted by whole tissues, essential oils, extracts, or individual compounds now encompasses ticks, fleas, termites, ants, mosquitoes, bacteria, a water mold, fungi and browsing animals. Nootkatone, is a heartwood component with sufficient activity against arthropods to warrant research focused toward potential development as a commercial repellent and biopesticide for ticks, mosquitoes and possibly other arthropods that vector human and animal pathogens.     

Chemical Cues that Guide Female Reproduction in <Emphasis Type="Italic">Drosophila melanogaster</Emphasis>

Chemicals released into the environment by food, predators and conspecifics play critical roles in Drosophila reproduction. Females and males live in an environment full of smells, whose molecules communicate to them the availability of food, potential mates, competitors or predators. Volatile chemicals derived from fruit, yeast growing on the fruit, and flies already present on the fruit attract Drosophila, concentrating flies at food sites, where they will also mate. Species-specific cuticular hydrocarbons displayed on female Drosophila as they mature are sensed by males and act as pheromones to stimulate mating by conspecific males and inhibit heterospecific mating. The pheromonal profile of a female is also responsive to her nutritional environment, providing an honest signal of her fertility potential. After mating, cuticular and semen hydrocarbons transferred by the male change the female’s chemical profile. These molecules make the female less attractive to other males, thus protecting her mate’s sperm investment. Females have evolved the capacity to counteract this inhibition by ejecting the semen hydrocarbon (along with the rest of the remaining ejaculate) a few hours after mating. Although this ejection can temporarily restore the female’s attractiveness, shortly thereafter another male pheromone, a seminal peptide, decreases the female’s propensity to re-mate, thus continuing to protect the male’s investment. Females use olfaction and taste sensing to select optimal egg-laying sites, integrating cues for the availability of food for her offspring, and the presence of other flies and of harmful species. We argue that taking into account evolutionary considerations such as sexual conflict, and the ecological conditions in which flies live, is helpful in understanding the role of highly species-specific pheromones and blends thereof, as well as an individual’s response to the chemical cues in its environment.     

Isolation and Identification of Host Cues from Mango, <Emphasis Type="Italic">Mangifera indica</Emphasis>, That Attract Gravid Female Oriental Fruit fly, <Emphasis Type="Italic">Bactrocera dorsalis</Emphasis>

The oriental fruit fly, Bactrocera dorsalis, is an economically damaging, polyphagous pest of fruit crops in South-East Asia and Hawaii, and a quarantine pest in other parts of the world. The objective of our study was to identify new attractants for B. dorsalis from overripe mango fruits. Headspace samples of volatiles were collected from two cultivars of mango, ‘Alphonso’ and ‘Chausa’, and a strong positive behavioral response was observed when female B. dorsalis were exposed to these volatiles in olfactometer bioassays. Coupled GC-EAG with female B. dorsalis revealed 7 compounds from ‘Alphonso’ headspace and 15 compounds from ‘Chausa’ headspace that elicited an EAG response. The EAG-active compounds, from ‘Alphonso’, were identified, using GC-MS, as heptane, myrcene, (Z)-ocimene, (E)-ocimene, allo-ocimene, (Z)-myroxide, and γ-octalactone, with the two ocimene isomers being the dominant compounds. The EAG-active compounds from ‘Chausa’ were 3-hydroxy-2-butanone, 3-methyl-1-butanol, ethyl butanoate, ethyl methacrylate, ethyl crotonate, ethyl tiglate, 1-octen-3-ol, ethyl hexanoate, 3-carene, p-cymene, ethyl sorbate, α-terpinolene, phenyl ethyl alcohol, ethyl octanoate, and benzothiazole. Individual compounds were significantly attractive when a standard dose (1 μg on filter paper) was tested in the olfactometer. Furthermore, synthetic blends with the same concentration and ratio of compounds as in the natural headspace samples were highly attractive (P < 0.001), and in a choice test, fruit flies did not show any preference for the natural samples over the synthetic blends. Results are discussed in relation to developing a lure for female B. dorsalis to bait traps with.     

Non-Volatile Intact Indole Glucosinolates are Host Recognition Cues for Ovipositing <Emphasis Type="Italic">Plutella xylostella</Emphasis>

The diamondback moth (Plutella xylostella), a crucifer-specialist pest, has been documented to employ glucosinolates as host recognition cues for oviposition. Through the use of mutant Arabidopsis thaliana plants, we investigated the role of specific classes of glucosinolates in the signaling of oviposition by P. xylostella in vivo. Indole glucosinolate production in A. thaliana was found to be crucial in attracting oviposition. Additionally, indole glucosinolates functioned as oviposition cues only when in their intact form. 4-Methoxy-indol-3-ylmethylglucosinolate was implicated as an especially strong oviposition attractant in vitro, suggesting that indole glucosinolate secondary structure may play a role in P. xylostella host recognition as well. Aliphatic glucosinolate-derived breakdown products were found to attract P. xylostella, but only after damage or in the absence of indole glucosinolates. Furthermore, mutant plants lacking both intact indole glucosinolates and aliphatic glucosinolate breakdown products exhibited decreased oviposition attractiveness beyond that of the progenitor mutants lacking either component of the glucosinolate-myrosinase system. Therefore, we conclude that nonvolatile indole glucosinolates and volatile aliphatic glucosinolate breakdown products both appear to play important roles as host recognition cues for P. xylostella oviposition.     

Semiochemicals used in Host Location by the Coffee berry Borer, <Emphasis Type="Italic">Hypothenemus hampei</Emphasis>

The coffee berry borer, Hypothenemus hampei is a serious pest in many coffee growing countries. Electrophysiological and behavioral responses of H. hampei to volatiles of different phenological stages of coffee, Coffea arabica, fruits were studied in order to identify volatile semiochemicals used in host location. Volatiles were collected from different phenological stages of C. arabica fruit by air entrainment. Electrophysiological recordings were made from insect antennae. Behavioral assays were carried out using a Perspex four-arm olfactometer. Insects spent significantly more time in the region of the olfactometer where ripe and dry fruit volatiles were present compared to control regions. Coupled gas chromatography—electroantennography revealed the presence of six electrophysiologically active compounds in C. arabica volatiles. These were identified by using GC and GC-MS as methylcyclohexane, ethylbenzene, nonane, 1-octen-3-ol, (R)-limonene, and (R)-3-ethyl-4-methylpentanol. In the olfactometer bioassay, H. hampei showed a significant response to 3-ethyl-4-methylpentanol, methylcyclohexane, nonane, ethylbenzene, and a synthetic blend of these four compounds. Attraction to the synthetic blend was comparable to that for the natural sample. The significance of the study is discussed in terms of semiochemical based pest management methods of the coffee berry borer.     

Oxidation kinetics for <Emphasis Type="Italic">cis</Emphasis>-9,<Emphasis Type="Italic">trans</Emphasis>-11 and <Emphasis Type="Italic">trans</Emphasis>-10,<Emphasis Type="Italic">cis</Emphasis>-12 isomers of CLA

The autoxidation processes of the cis-9,trans-11 (c9,t11) and trans-10,cis-12 (t10,c12) isomers of CLA were separately observed at ca. 0% RH and different temperatures. The t10,c12 CLA oxidized faster than the c9,t11 isomer at all tested temperatures. The first half of the oxidation process of t10,c12 CLA obeyed an autocatalytic-type rate expression, but the latter half followed first-order kinetics. On the other hand, the entire oxidation process of c9,t11 CLA could be expressed by the autocatalytic-type rate expression. The apparent activation energies and frequency factors for the autoxidation of the isomers were estimated from the rate constants obtained at various temperatures based on the Arrhenius equation. The apparent activation energies for the CLA isomers were greater than those for the nonconjugated n−6 and n−3 PUFA or their esters. However, the enthalpyentropy compensation held during the autoxidation of both the CLA and PUFA. This suggested that the autoxidation mechanisms for the CLA and PUFA were essentially the same.     

Volatile and Contact Chemical Cues Associated with Host and Mate Recognition Behavior of <Emphasis Type="Italic">Sphenophorus venatus</Emphasis> and <Emphasis Type="Italic">Sphenophorus parvulus</Emphasis> (Coleoptera: Dryophthoridae)

Beetles in the genus Sphenophorus Schönherr, or billbugs, potentially utilize both volatile and non-volatile behavior-modifying chemical signals. These insects are widely distributed across North America, often occurring in multi-species assemblages in grasses. However, details about their host- and mate-finding behavior are poorly understood. This study tested the hypothesis that volatile organic compounds from host-plants and conspecifics direct the dispersal behavior of hunting billbug S. venatus Say. Further, we characterized the cuticular hydrocarbon profiles of two widespread pest species, S. venatus and bluegrass billbug S. parvulus Gyllenhaal, to assess the potential role of contact pheromones in mate-recognition. In Y-tube olfactometer bioassays, S. venatus males were attracted to a combination of conspecifics and Cynodon dactylon host-plant material, as well as C. dactylon plant material alone. S. venatus females were attracted to a combination of male conspecifics and host-plants but were also attracted to male conspecifics alone. Field evaluation of a putative male-produced aggregation pheromone, 2-methyl-4-octanol, identified from two congeners, S. levis Vaurie and S. incurrens Gyllenhaal, did not support the hypothesis that S. venatus and S. parvulus were also attracted to this compound. Gas chromatography-mass spectrometry analysis of S. venatus and S. parvulus whole-body cuticular extracts indicated a series of hydrocarbons with qualitative and quantitative interspecific variation in addition to intraspecific quantitative variation between males and females. This study provides the first evidence that S. venatus orients toward host- and insect-derived volatile organic compounds and substantiates the presence of species-specific cuticular hydrocarbons that could serve as contact pheromones for sympatric Sphenophorus species.     

Caterpillar Footprints as Host Location Kairomones for <Emphasis Type="Italic">Cotesia marginiventris</Emphasis>: Persistence and Chemical Nature

Herbivores walking over the epicuticular wax layer of a plant may leave tracks that disclose their presence to hunting predators or parasitoids. The braconid wasp Cotesia marginiventris is a solitary parasitoid of young noctuid caterpillars. It can locate potential hosts from a distance by orienting toward the scent of herbivore-damaged plants. Upon landing on the caterpillars’ food plant, the female parasitoid searches for further cues (kairomones) that confirm the presence of a suitable host. In a previous study, we showed that C. marginiventris recognizes the chemical footprints of absent Spodoptera frugiperda caterpillars on a leaf. Here, we report on the persistence and chemical nature of this host location kairomone. In a series of behavioral assays, we confirmed that caterpillars of S. frugiperda leave chemical tracks that elicit characteristic antennation behavior in C. marginiventris for up to 2 days. Both hexane extracts of caterpillar footprints and of the larvae’s ventral cuticle induced antennation and contained almost identical long-chain hydrocarbons, thus suggesting the prolegs and claspers as the kairomones’ main source. A series of linear C₂₁ to C₃₂ alkanes accounted for ca 90% of all identified compounds. Female wasps showed significant antennation responses on leaves treated with a reconstructed blend of these n-alkanes. However, wasp responses were relatively weak. Therefore, we presume that minor compounds, such as monomethyl-branched alkanes, which were also found, may contribute additionally to host recognition.     

Chemotaxis of the Pinewood Nematode, <Emphasis Type="Italic">Bursaphelenchus xylophilus</Emphasis>, to Volatiles Associated with Host Pine, <Emphasis Type="Italic">Pinus massoniana</Emphasis>, and its Vector <Emphasis Type="Italic">Monochamus alternatus</Emphasis>

The pinewood nematode (PWN), Bursaphelenchus xylophilus, the most important invasive species in pine forests of Asia, is transported to new pine hosts by beetles of the genus Monochamus. Third-stage dispersal juveniles (J_III) aggregate in pupal chambers around the vector as it matures. We demonstrated that the ratio of three terpenes (α-pinene, β-pinene, and longifolene at 1:2.7:1.1) released by larval Monochamus alternatus strongly attract J_III, whereas the different ratio (1:0.1:0.01) of these three terpenes found in healthy xylem of Pinus massoniana attracts only the propagative stage (J_n) of the nematode. We suggest that the volatiles produced by the host plants could be the basis of a chemoecological relationship between plant parasitic nematodes and their vector insects. Capture of J_III with terpene-baited trap tubes deployed for 2 hr in the field was demonstrated. This technique may lead to the development of rapid sampling methodologies for use at either ports-of-entry or in the field.     

Comparison of Postprandial Oleic Acid, 9<Emphasis Type="Italic">c,</Emphasis>11<Emphasis Type="Italic">t</Emphasis> CLA and 10<Emphasis Type="Italic">t,</Emphasis>12<Emphasis Type="Italic">c</Emphasis> CLA Oxidation in Healthy Moderately Overweight Subjects

Few studies report the individual effect of 9c,11t- and 10t,12c-CLA on human energy metabolism. We compared the postprandial oxidative metabolism of 9c,11t- and 10t,12c-CLA and oleic acid (9c-18:1) in 22 healthy moderately overweight volunteers. After 24 weeks supplementation with 9c,11t-, 10t,12c-CLA or 9c-18:1 (3 g/day), subjects consumed a single oral bolus of the appropriate [1-¹³C]-labeled fatty acid. 8 h post-dose, cumulative oxidation was similar for 9c-18:1 and 10t,12c (P = 0.66), but significantly higher for 9c,11t (P < 0.01).     

(1<Emphasis Type="Italic">S</Emphasis>,3<Emphasis Type="Italic">R</Emphasis>)-<Emphasis Type="Italic">cis</Emphasis>-Chrysanthemyl Tiglate: Sex Pheromone of the Striped Mealybug, <Emphasis Type="Italic">Ferrisia virgata</Emphasis>

Derivatives of 2,2-dimethyl-3-(2-methylprop-1-enyl)cyclopropanecarboxylic acid (chrysanthemic acid) are classic natural pyrethroids discovered in pyrethrum plants and show insecticidal activity. Chrysanthemic acid, with two asymmetric carbons, has four possible stereoisomers, and most natural pyrethroids have the (1R,3R)-trans configuration. Interestingly, chrysanthemic acid–related structures are also found in insect sex pheromones; carboxylic esters of (1R,3R)-trans-(2,2-dimethyl-3-(2-methylprop-1-enyl)cyclopropyl)methanol (chrysanthemyl alcohol) have been reported from two mealybug species. In the present study, another ester of chrysanthemyl alcohol was discovered from the striped mealybug, Ferrisia virgata (Cockerell), as its pheromone. By means of gas chromatography–mass spectrometry, nuclear magnetic resonance spectrometry, and high-performance liquid chromatography analyses using a chiral stationary phase column and authentic standards, the pheromone was identified as (1S,3R)-(?)-cis-chrysanthemyl tiglate. The (1S,3R)-enantiomer strongly attracted adult males in a greenhouse trapping bioassay, whereas the other enantiomers showed only weak activity. The cis configuration of the chrysanthemic acid–related structure appears to be relatively scarce in nature, and this is the first example reported from arthropods.     

Specificity in Chemical Profiles of Workers,Brood and Mutualistic Fungi in <Emphasis Type="Italic">Atta</Emphasis>, <Emphasis Type="Italic">Acromyrmex,</Emphasis> and <Emphasis Type="Italic">Sericomyrmex</Emphasis> Fungus-growing Ants

Neotropical attine ants live in obligatory symbiosis with a fungus that they grow for food on a substrate of primarily plant material harvested by workers. Nestmate recognition is likely based on chemical cues as in most other social insects, but recent studies have indicated that both the ants and their mutualistic fungi may contribute to the recognition templates. To investigate the within-colony variation in chemical profiles, we extracted and identified compounds from the cuticle of workers, the postpharyngeal gland of workers, ant pupae and larvae, and the fungal symbiont of three species of higher attine ants: Atta colombica, Acromyrmex echinatior, and Sericomyrmex amabilis. The relative proportions of identified compounds were compared and represented 11 classes: n-alkanes, alkenes, branched methylalkanes, branched dimethylalkanes, trimethylalkanes, branched alkenes, aldehydes, alcohols, acetates, acids, and esters. The chemical profiles in all three species are likely to be sufficiently different to allow discrimination at the species and colony level and sufficiently similar within colonies to generate a relatively constant colony-specific chemical gestalt. The relative likelihood of individual compounds being derived from the ants, the ant brood, or the fungal symbiont are discussed. We hypothesize that hydrocarbons are particularly important as recognition cues because they appear to simultaneously allow the assessment of developmental stages and the identification of symbiont, colony, and species.     

<Emphasis Type="Italic">Santalum </Emphasis><Emphasis Type="Italic">insulare</Emphasis> Acetylenic Fatty Acid Seed Oils: Comparison within the <Emphasis Type="Italic">Santalum</Emphasis> Genus

The sandalwood kernels of Santalum insulare (Santalaceae) collected in French Polynesia give seed oils containing significant amounts of ximenynic acid, E-11-octadecen-9-oic acid (64–86%). Fatty acid (FA) identifications were performed by gas chromatography/mass spectrometry (GC/MS) of FA methyl esters. Among the other main eight identified fatty acids, oleic acid was found at a 7–28% level. The content in stearolic acid, octadec-9-ynoic acid, was low (0.7–3.0%). An inverse relationship was demonstrated between ximenynic acid and oleic acid using 20 seed oils. Results obtained have been compared to other previously published data on species belonging to the Santalum genus, using multivariate statistical analysis. The relative FA S. insulare composition, rich in ximenynic acid is in the same order of those given for S. album or S. obtusifolium. The other compared species (S. acuminatum, S. lanceolatum, S. spicatum and S. murrayanum) are richer in oleic acid (40–59%) with some little differences in linolenic content.     

Chemical Composition and Nutritional Characteristics of the Seed Oil of Wild <Emphasis Type="Italic">Lathyrus</Emphasis>, <Emphasis Type="Italic">Lens</Emphasis> and <Emphasis Type="Italic">Pisum</Emphasis> Species from Southern Spain

The fatty acid composition of the seed oil of 19 wild legume species from southern Spain was analyzed by gas chromatography. The main seed oil fatty acids ranged from C_14:0 to C_20:0. Among unsaturated fatty acids, the most abundant were linoleic, oleic and linolenic acids, except for Lathyrus angulatus, L. aphaca, L. clymenum, L. sphaericus and L. nigricans where C_18:3 contents were higher than C_18:1 contents. Palmitic acid was the most abundant saturated acid in studied species, ranging from 11.6% in Lathyrus sativus to 19.3% in Lens nigricans. All studied species showed higher amounts of total unsaturated fatty acids than saturated ones. Among studied species, the ω6/ω3 ratio was variable, ranging from 2.0% in L. nigricans to 13.8% in L. sativus, there being eight species in which the ω6/ω3 ratio was below 5. The fatty acids observed in these plants supports the use of these plants as a source of important dietary lipids.     

Sexual Deception in the <Emphasis Type="Italic">Eucera</Emphasis>-Pollinated <Emphasis Type="Italic">Ophrys leochroma</Emphasis>: A Chemical Intermediate between Wasp- and <Emphasis Type="Italic">Andrena-</Emphasis>Pollinated Species

Ophrys flowers mimic sex pheromones of attractive females of their pollinators and attract males, which attempt to copulate with the flower and thereby pollinate it. Virgin females and orchid flowers are known to use the same chemical compounds in order to attract males. The composition of the sex pheromone and its floral analogue, however, vary between pollinator genera. Wasp-pollinated Ophrys species attract their pollinators by using polar hydroxy acids, whereas Andrena-pollinated species use a mixture of non-polar hydrocarbons. The phylogeny of Ophrys shows that its evolution was marked by episodes of rapid diversification coinciding with shifts to different pollinator groups: from wasps to Eucera and consequently to Andrena and other bees. To gain further insights, we studied pollinator attraction in O. leochroma in the context of intra- and inter-generic pollinator shifts, radiation, and diversification in the genus Ophrys. Our model species, O. leochroma, is pollinated by Eucera kullenbergi males and lies in the phylogeny between the wasp and Andrena-pollinated species; therefore, it is a remarkable point to understand pollinator shifts. We collected surface extracts of attractive E. kullenbergi females and labellum extracts of O. leochroma and analyzed them by using gas chromatography with electroantennographic detection (GC-EAD) and gas chromatography coupled with mass spectrometry (GC-MS). We also performed field bioassays. Our results show that O. leochroma mimics the sex pheromone of its pollinator’s female by using aldehydes, alcohols, fatty acids, and non-polar compounds (hydrocarbons). Therefore, in terms of the chemistry of pollinator attraction, Eucera-pollinated Ophrys species might represent an intermediate stage between wasp- and Andrena-pollinated orchid species.     

Olfactory Preferences of the Parasitic Nematode <Emphasis Type="Italic">Howardula aoronymphium</Emphasis> and its Insect Host <Emphasis Type="Italic">Drosophila falleni</Emphasis>

Many parasitic nematodes have an environmental infective stage that searches for hosts. Olfaction plays an important role in this process, with nematodes navigating their environment using host-emitted and environmental olfactory cues. The interactions between parasitic nematodes and their hosts are also influenced by the olfactory behaviors of the host, since host olfactory preferences drive behaviors that may facilitate or impede parasitic infection. However, how olfaction shapes parasite-host interactions is poorly understood. Here we investigated this question using the insect-parasitic nematode Howardula aoronymphium and its host, the mushroom fly Drosophila falleni. We found that both H. aoronymphium and D. falleni are attracted to mushroom odor and a subset of mushroom-derived odorants, but they have divergent olfactory preferences that are tuned to different mushroom odorants despite their shared mushroom environment. H. aoronymphium and D. falleni respond more narrowly to odorants than Caenorhabditis elegans and Drosophila melanogaster, consistent with their more specialized niches. Infection of D. falleni with H. aoronymphium alters its olfactory preferences, rendering it more narrowly tuned to mushroom odor. Our results establish H. aoronymphium-D. falleni as a model system for studying olfaction in the context of parasite-host interactions.