EAG Responses Research Articles

Electrophysiological and behavioural responses of Melolontha melolontha to saturated and unsaturated aliphatic alcohols

AbstractMale Melolontha cockchafers are known to use green leaf volatiles induced by female feeding on host plants for their mate location. Earlier studies of the response of the European cockchafer, Melolontha melolontha L. (Coleoptera: Scarabaeidae), to different green leaf aldehydes, alcohols, and acetates revealed that only green leaf alcohols were attractive to males in the field. Females were not attracted at all by these volatiles. Here, we present a study that aimed to elucidate the structure–activity relationships of aliphatic alcohols. Both behavioural and physiological responses were studied in male and female M. melolontha by field tests and electroantennography. The compounds tested were saturated aliphatic alcohols with chain lengths between five and eight carbon atoms. Furthermore, the cockchafer's response to six‐carbon alcohols with (E)‐2‐, (E)‐3‐, (Z)‐2‐, (Z)‐3‐, and (Z)‐4‐configurated double bonds was tested. All compounds elicited dose‐dependent responses on the antennae of both sexes. In general, males showed a stronger normalized EAG response to the stimuli than females. However, only the naturally occurring six‐carbon alcohols, i.e., 1‐hexanol (E)‐2‐, (Z)‐3, and (E)‐3‐hexen‐1‐ol were attractive to M. melolontha males in the field. Females were not attracted to any of the tested compounds, confirming previous results on the olfactory orientation of Melolontha cockchafers.

The role of volatile semiochemicals in mediating host location and selection by nuisance and disease‐transmitting cattle flies

The role of volatile semiochemicals in mediating the location and selection within herds of Holstein-Friesian heifers by nuisance and disease-transmitting cattle flies was investigated using coupled gas chromatography-electrophysiology (GC-EAG), coupled gas chromatography-mass spectrometry (GC-MS), electrophysiology (EAG), laboratory behaviour and field studies. Using volatile extracts collected by air entrainment from heifers in the Netherlands, a number of active peaks were located by coupled GC-EAG for Musca autumnalis (de Geer) (Diptera: Muscidae) and Haematobia irritans (L.) (Diptera: Muscidae). Volatile samples were also collected from two heifers in Denmark shown in previous counting experiments to differ significantly in their fly loads. Coupled GC-EAG using Ha. irritans antennae revealed differences in the EAG response to the samples, with additional EAG activity in the sample collected from the heifer with the lower fly load. To identify more EAG active compounds, volatiles were also collected from 48-h-old urine by air entrainment. In total, 23 compounds were located and identified by coupled GC-EAG and GC-MS. Further electrophysiological testing of these compounds with five fly species [M. autumnalis, Ha. irritans, Hydrotaea irritans (L.) (Diptera: Muscidae), Stomoxys calcitrans (L.) (Diptera: Musicidae) and Wohlfahrtia magnifica (Schiner) (Diptera: Sarcophagidae)] showed that only some of the compounds were physiologically active across the range of flies tested. These included 1-octen-3-ol, 6-methyl-5-hepten-2-one, (Z)-3-hexen-1-ol, naphthalene, and all EAG active compounds identified from urine. Compounds showing significant EAG activity were tested for behavioural activity using a wind-tunnel designed for measuring upwind flight behaviour. At certain concentrations, 1-octen-3-ol, 6-methyl-5-hepten-2-one and 3-octanol increased upwind flight, whereas naphthalene, propyl butanoate and linalool reduced upwind flight. In field studies using small herds of heifers ranked according to their fly load, individual slow-release formulations of 1-octen-3-ol and 6-methyl-5-hepten-2-one, when applied to low and high fly loading heifers, reduced fly loads on these individuals. This study provides evidence for the hypothesis that the natural differential attractiveness within herds of Holstein-Freisian heifers, i.e. a single host species, for cattle flies is partly due to differences in volatile semiochemicals emitted from the host. It is suggested that this phenomenon applies to other vertebrate host species and their associated insect pests.

Behavioural and electrophysiological responses of the malaria mosquito Anopheles gambiae Giles sensu stricto (Diptera: Culicidae) to human skin emanations

Behavioural and electrophysiological responses of Anopheles gambiae Giles sensu stricto (Diptera: Culicidae) to human skin emanations collected on glass beads were studied using a dual-port olfactometer and electroantannography. Glass beads to which skin emanations from human hands had been transferred elicited a level of attraction similar to a human hand. The attractiveness of these handled glass beads faded away 4 h after transfer onto the beads. Storage at -20 degrees C for up to 8 weeks showed a decreased but still attractive effect of the beads. In a choice test between one individual and four others, the emanations from the reference individual were significantly more attractive in three out of four cases. The headspace of handled glass beads elicited a dose-dependent EAG response. The substances causing EAG activity could be removed partially by dichloromethane, ethanol and pentane-ether. Glass beads provide a suitable neutral substrate for the transfer of human odour to enable chemical analysis of the human skin emanations for identification of kairomones of anthropophilic mosquitoes.

Geographic Variation in Pheromone Chemistry, Antennal Electrophysiology, and Pheromone-Mediated Trap Catch of North American Populations of the Obliquebanded Leafroller

The total and relative amounts of (Z)-11-tetradecenyl acetate (Z11-14:Ac), (E)-11-tetradecenyl acetate (E11-14:Ac), (Z)-11-tetradecen-1-ol (Z11-14:OH) and (Z)-11-tetradecenal (Z11-14:Al), and the EAG response of male antennae to these pheromone gland compounds were compared in laboratory reared Choristoneura rosaceana Harris (Lepidoptera: Tortricidae) from British Columbia, Michigan, Ontario, New York, and Quebec. A field trapping experiment was conducted in each of these locations to determine the effect of Z11-14:Al on the numbers of moths captured. The amount of each of the four pheromone-gland compounds declined successively in moths from British Columbia, Quebec, Ontario, Michigan, and New York. The relative amount of Z11-14:Ac was greatest in moths from New York and smallest in moths from Ontario, whereas the relative amount of E11-14:Ac was greatest in moths from Ontario and smallest in moths from British Columbia. Moths from Ontario, Quebec, British Columbia, Michigan, and New York contained decreasing relative amounts of Z11-14:OH and Z11-14:Al. There was a trend of increasing antennal sensitivity to each of the four pheromone-gland compounds in moths from New York, Michigan, Ontario, Quebec, and British Columbia. The addition of 1% Z11:Al to a three compound blend of Z11-14:Ac, E11-14:Ac and Z11-14:OH (97:2:1) resulted in a >twofold increase in average trap catch in British Columbia, Ontario, and Quebec; this compound had no effect on trap catch in Michigan or New York.

Mate location in the green capsid bug, Lygocoris pabulinus

AbstractHeadspace extracts from female Lygocoris pabulinus (L.) (Heteroptera: Miridae) as well as female leg extracts attracted male L. pabulinus in Y‐track olfactometer bioassays. In contrast, only female leg extracts were active in the vibration bioassay. Male extracts had no activity at all in either bioassay. When the female leg extract was analysed by coupled gas chromatography‐electroantennography (Z)‐9‐pentacosene and (Z)‐7‐pentacosene were EAD‐active. (Z)‐9‐Pentacosene and sometimes (Z)‐7‐pentacosene were also observed in female headspace extracts. EAG responses could be obtained for (Z)‐9‐pentacosene. A mixture of (Z)‐9‐pentacosene and (Z)‐7‐pentacosene in the ratio 5 : 1 elicited vibration behaviour in males. The results indicate that these alkenes are important cues for male L. pabulinus in mate location behaviour.

Minor components in the sex pheromone of legume podborer: Maruca vitrata development of an attractive blend.

The legume podborer, Maruca vitrata (syn. M. testulalis) (F.) (Lepidoptera: Pyralidae) is a pantropical pest of legume crops. Sex pheromone was collected by gland extraction or trapping of volatiles from virgin female moths originating in India, West Africa, or Taiwan. Analysis by GC-EAG and GC-MS confirmed previously published findings that (E,E)-10,12-hexadecadienal is the most abundant EAG-active component with 2-5% of (E,E)-10,12-hexadecadienol also present. At least one other EAG response was detected at retention times typical of monounsaturated hexadecenals or tetradecenyl acetates, but neither could be detected by GC-MS. Laboratory wind-tunnel bioassays and a field bioassay of blends of (E,E)-10,12-hexadecadienal with (E,E )-10,12-hexadecadienol and a range of monounsaturated hexadecenal and tetradecenyl acetate isomers indicated greatest attraction of males was to those including (E,E)-10,12-hexadecadienol and (E)-10-hexadecenal as minor components. In subsequent trapping experiments in cowpea fields in Benin, traps baited with a three-component blend of (E,E)-10,12-hexadecadienal and these two minor components in a 100:5:5 ratio caught significantly more males than traps baited with the major component alone, either two-component blend, or virgin female moths. Further blend optimization experiments did not produce a more attractive blend. No significant differences in catches were found between traps baited with polyethylene vials or rubber septa, or between lures containing 0.01 and 0.1 mg of synthetic pheromone. Significant numbers of female M. vitrata moths, up to 50% of total catches, were trapped with synthetic blends but not with virgin females. At present there is no clear explanation for this almost unprecedented finding, but the phenomenon may improve the predictive power of traps for population monitoring.

Electroantennographic and behavioral responses of the sphinx moth Manduca sexta to host plant headspace volatiles.

Coupled gas chromatography with electroantennographic detection (GC-EAD) using antennae of adult female Manduca sexta was employed to screen for olfactory stimulants present in headspace collections from four species of larval host plants belonging to two families: Solanaceae--Lycopersicon esculentum (tomato), Capiscum annuum (bell pepper), and Datura wrightii; and Martyniaceae--Pronboscideaparviflora. Headspace volatiles were collected from undamaged foliage of potted, living plants. GC-EAD revealed 23 EAD-active compounds, of which 15 were identified by GC-mass spectrometry. Identified compounds included aliphatic, aromatic, and terpenoid compounds bearing a range of functional groups. Nine EAD-active compounds were common to all four host plant species: (Z)-3-hexenyl acetate, nonanal, decanal, phenylacetaldehyde, methyl salicylate, benzyl alcohol, geranyl acetone, (E)-nerolidol, and one unidentified compound. Behavioral responses of female moths to an eight-component synthetic blend of selected tomato headspace volatiles were tested in a laboratory wind tunnel. Females were attracted to the blend. A comparison of responses from antennae of males and females to bell pepper headspace volatiles revealed that males responded to the same suite of volatiles as females, except for (Z)-3-hexenyl benzoate. EAD responses of males also were lower for (Z)-and (E)-nerolidol and one unidentified compound. Electroantennogram EAG dose-response curves for the 15 identified EAD-active volatiles were recorded. At the higher test doses (10-100 microg), female antennae yielded larger EAG responses to terpenoids and to aliphatic and aromatic esters. Male antennae did respond to the higher doses of (Z)-3-hexenyl benzoate, indicating that they can detect this compound. On the basis of ubiquity of the EAD-active volatiles identified to date in host plant headspace collections, we suggest that M. sexta uses a suite of volatiles to locate and identify appropriate host plants.

Identification of odors from overripe mango that attract vinegar flies, Drosophila melanogaster.

Bioassays with a variety of overripe fruits, including mango, plum, pear, and grape, and their extracts showed that odors from overripe mango were most attractive to adult vinegar flies, Drosophila melanogaster. Combined gas chromatography-electroantennographic detection (GC-EAD) analyses of solid-phase microextraction (SPME) and Tenax extracts of overripe mango odors showed that several volatile compounds, including ethanol, acetic acid, amyl acetate, 2-phenylethanol, and phenylethyl acetate elicited significant EAG responses from antennae of female flies. Most of the volatile compounds in the extracts were identified by mass spectral and retention index comparisons with synthetic standards. In cage bioassays, lures with a blend of ethanol, acetic acid, and 2-phenylethanol in a ratio of 1:22:5 attracted six times more flies than any single EAG-active compound. This blend also attracted four times more flies than traps baited with overripe mango or unripe mango. However, in field trials, the blend was not as attractive as suggested by the laboratory bioassay.

The role of plant odours in the leafminer Liriomyza sativae (Diptera: Agromyzidae) and its parasitoid Diglyphus isaea (Hymenoptera: Eulophidae): Orientation towards the host habitat

Electroantennogram responses of the polyphagous leafminer Liriomyza sativae and its generalist parasitoid Diglyphus isaea to host and non-host plant odours of L. sativae were investigated. The odours of healthy leaves can elicit distinct EAG responses in L. sativae. The EAG responses to the odours of the host plants, bean and tomato, were stronger than to non-host plants, Chinese rose and morning glory. Neither healthy host nor non-host plants of the leafminer elicited distinctive EAG responses in the parasitoid, D. isaea. Odours of physically damaged leaves, no matter whether of host or non-host plants, increased strongly the EAG responses of the leafminer and its parasitoid. We compared the EAG responses of D. isaea to bean leaves of different status. The odour of mined leaves elicited distinct EAG responses, which were weaker than those of physically damaged ones. No differences were detected in the EAG response to leaves with empty mines, healthy leaves or an air control. The role of plant odours in host location of the leafminer and its parasitoid is discussed.

GC-EAG-analysis of volatiles from Brussels sprouts plants damaged by two species of Pieris caterpillars: olfactory receptive range of a specialist and a generalist parasitoid wasp species

Feeding by Pieris brassicae or P. rapae caterpillars on Brussels sprouts plants induces the emission of synomones that attract natural enemies of the caterpillars, Cotesia glomerata, a generalist parasitoid, and C. rubecula, a specialist on P. rapae. Previous research on this tritrophic system has identified a large number of volatiles in the headspace of herbivore-damaged Brussels sprouts plants, and this paper addresses the question which of these volatiles are perceived by the two parasitoid species. Headspace odors from both P. brassicae- and P. rapae-damaged Brussels sprouts plants were analyzed by coupled gas chromatography electro- antennogram (GC-EAG) detection. Twenty volatiles evoked consistent EAG reactions in the antennae of both species and nineteen of these volatiles could be identified with GC-MS. One component that could not be identified due to its low concentration, evoked EAG responses in antennae of C. rubecula only. Possible consequences for searching behavior of the two parasitoid species are discussed.

Role of plant volatiles in host plant location of the leafminer, Liriomyza sativae (Diptera: Agromyzidae)

Abstract The role of plant volatiles in host plant location of the leafminer Liriomyza sativae Blanchard was studied. Four types of antennal sensilla were identified on the funiculus by scanning electron microscopy: trichoid, basiconic, clavate and grooved sensilla. An olfactory pit, containing groups of sensilla, was present on the ventral side of the funiculus. No sexual difference was detected in sensilla diversity and distribution. In behavioural assays, both males and females were attracted by the odour of the bean Phaseolus vulgaris L. They had distinct EAG responses to the bean odour. No significant sexual difference was found in behaviour or EAG responses.Electroantennograms were recorded from female L. sativae to 14 plant volatile compounds. The most distinct EAG responses were obtained for: (1) the general green leaf volatiles 1‐hexanol (E)‐2‐hexen‐1‐ol, (E)‐3‐hexen‐1‐ol and its isomers, (Z)‐3‐hexen‐1‐ol, the acetate (E)‐3‐hexenylacetate and the aldehyde hexanal; and (2) limonene, a compound associated with tomato, which is a key host plant of this insect. Other volatile compounds associated with host plants, such as α‐pinene, myrcene, β‐caryophyllene, and eugenol did not elicit responses. The ability of this insect to locate a host plant appears to be augmented by the perception of a combination of host‐specific and general green leaf volatiles. A modification of the EAG recording method of Dipteran species was provided.

Electroantennogram responses of Apanteles obliquae (Hym., Braconidae) to various infochemicals

Abstract: The electroantennogram recording technique (EAG) was used to study the olfactory sensitivity of Apanteles obliquae (Hym., Braconidae), a gregarious larval endoparasitoid of Spilosoma obliqua (Walker) (Lep., Arctiidae), to 25 general plant volatiles belonging to alcohol, aldehyde and terpenoid groups and also to volatiles from the host and plant–host complex. The EAG data indicated different olfactory sensitivity between the sexes, not only to individual plant volatiles but also to the volatiles from host and plant–host complex. Females were found to be more responsive than males. However the synthetic sex pheromone blend of the host insect elicited similar EAG responses in both sexes. The EAG data of the present study is correlated with the reported behaviour observed in other parasitoids.

Attractant volatiles released by female and male Triatoma infestans (Hemiptera: Reduviidae), a vector of chagas disease: chemical analysis and behavioral bioassay.

Volatiles emitted by male and female T infestans before and during copula were collected on Porapak-Q filters, desorbed with dichloromethane, and analyzed by gas chromotography and gas chromatography-mass spectrometry after confirmation of attractiveness in an arena bioassay. Chemical analysis confirmed the presence of (R,S) -2- and 3-methylbutan-1-ol in a 2:1 ratio; short chain acids (ethanoic to nonanoic acid); long chains acids decanoic to (Z)-9-octadecenoic acid; aliphatic aldehydes (hexanal to nonanal), benzaldehyde and dipropylsulphide from insects in copula. Electroantennographic studies conducted with a homologous series of aliphatic aldehydes on female and male T infestans showed that, for a given dose, EAG responses elicited from both sexes increased with increased chain length up to nonanal, after which EAG-activity declined. Attractiveness of non-acidic trace components identified in the volatiles were tested on male and female T. infestans, in an arena bioassay using a video tracking method. Aliphatic C6 to C10 aldehydes were tested: hexanal (1-100 microg) and heptanal (10 microg) were attractive to female T. infestans, high doses of octanal and nonanal (1-100 microg) were Unattractive to male and female T. infestans but low doses of nonanal (0.01-0.1 microg) were attractive to male T infestans. Benzaldehyde was highly attractive to female T. infestans at low doses (0.05- 0.1 microg). 3-methylbutan-1-ol was attractive to male T infestans at high dose (1,000 microg). (S) or (S,R) 2-methyl-butan-1-ol were attractive to males or females (1-1,000 microg). Blends of hexanal and benzaldehyde (20:1 and 40:1) showed an additive effect on attraction compared with hexanal alone, when tested on female T. infestans. The study has demonstrated the presence of a number of electrophysiologically and behaviorally active compounds in volatiles emitted by T. infestans in copula that may have a role in the postulated copulation pheromone.

Identification of odoriferous compounds from adults of a swallowtail butterfly, Papilio machaon (Lepidoptera: Papilionidae).

Adults, particularly males, of a papilionid butterfly, Papilio machaon hippocrates, emit a fairly strong scent perceivable by humans. We have identified a variety of volatile compounds (hydrocarbons, alcohols, aldehydes, ketones, esters, and so on) from the wings and bodies of both sexes of the butterfly. Male wings secreted n-dodecane, linalool and geranylacetone as major components together with small amounts of camphene, limonene, p-cymene, 2-phenylethanol, n-hexanal, n-decanal, isoamyl acetate, p-allylanisole, 2-pyrrolidone and other characteristic volatiles. The overall profile of volatile compounds detected from male body was quite different from that of the wings. Male body was devoid of camphene, 2-phenylethanol, n-hexanal but instead contained limonene, acetoin, a sesquiterpene hydrocarbon (C15H24, methyl n-octanoate, (E,E)-hepta-2,4-dienal, and another isomer of heptadienal as principal components, of which the last four compounds were specific to the body. All these substances seem to concurrently characterize the male odor. The chemical patterns of compounds found from female wings and body were essentially the same in quality as those of male wings and body, respectively, although their quantities in females were generally smaller than in males. Females, however, had a larger amount of acetamide than males. The chemical compositions of volatiles from the fore and hind wings of males were not greatly different from each other, and every component was considered to be present on all parts of the wings. This suggests that the scent-producing organs or scent-emitting pores are widely distributed on the whole wings. EAG responses of both sexes to 12 selected compounds identified from the butterfly were not strong at a dose of 1 microg, while both sexes showed relatively stronger responses to n-nonanal, methyl n-octanoate, D-limonene and linalool at a higher dose (10 microg). Although sexual difference in EAG response was not prominent, females appeared a little more sensitive, and n-nonanal and acetoin evoked significantly higher responses from females at 1 microg.

FLORAL SCENT OF Osmanthus fragrans DISCOURAGES FORAGING BEHAVIOR OF CABBAGE BUTTERFLY, Pieris rapae

Field observations revealed that flowers of Osmanthus fragrans attract few insect species despite their strong scent and vivid coloration (yellowish orange). Floral volatiles of the plant were thus examined for suspected repellency to Pieris rapae, a potential visitor that never visits this flower. The influence of odor components on the foraging behavior of adult butterflies was assessed by two behavioral bioassays: proboscis extension reflex (PER) in response to olfactory stimuli and the frequency of visits to artificial flowers. In the PER bioassays, a binary mixture composed of a test material and 2-phenylethanol (a positive standard compound) was used, and the negative effect of test materials was appraised based on the degree of suppression caused compared with the original PER performance evoked by 2-phenylethanol alone. Isopentane extract of the flower exhibited significant deterrency in the PER test and remarkable repellency in the flower-visiting experiment, thereby suggesting the presence of a repellent(s) in the floral volatiles. Fractionation of the extract by silica gel column chromatography yielded an active fraction (based on PER performance), which, upon GC and GC-MS analyses, was found to consist of γ-decalactone, β-ionone, and linalool oxide (LO) isomers (furanosides and pyranosides) as major components together with small amounts of linalool and α-ionone. γ-Decalactone was strongly deterrent in the PER test, and β-ionone and all LO isomers were weakly deterrent, while the other compounds exhibited no deterrency. In the flower-visiting tests, γ-decalactone showed notable repellency, whereas β-ionone had no repellent effect. Among the LO isomers, at least furanosides (cis/trans mixture) and cis-pyranoside were found to be weakly repellent. On the other hand, the butterfly showed strong to weak EAG responses to LO furanosides, cis-LO pyranoside, γ-decalactone, and β-ionone in decreasing order of intensity. The present results clearly indicate that the repellency of a compound is correlated with its deterrent effect on PER but not necessarily with antennal sensitivity.

Identification of a new lepidopteran sex pheromone in picogram quantities using an antennal biodetector: (8 E,10 Z)-tetradeca-8,10-dienal from Cameraria ohridella

A major sex attractant released by the virgin female of the horse-chestnut leafminer Cameraria ohridella Deschka et Dimić (Lepidoptera: Gracillariidae) which devastates horse-chestnut trees in Europe, was identified in picogram quantities as (8 E,10 Z)-tetradeca-8,10-dienal without using spectral methods. The identification solely relied on gas chromatography with electroantennographic detection (GC-EAD), calculation of Kovats' indices of the active principle on different GC phases, and construction of antennal response spectra (EAG response profiles) to C 12 and C 14 saturated and unsaturated standards with different functional groups. The dienal was prepared by a stereospecific synthesis and shown to be highly active for conspecific males in pg amounts and fully comparable to the natural substance.

Correction for differences in volatility among olfactory stimuli and effect on EAG responses of Dioryctria abietivorella to plant volatiles

We describe a method using paraffin oil solutions and gas chromatography to measure and correct for differences in volatility among test compounds applied to filter paper and to address the problem of minimizing solvent contributions to EAG responses. To examine the effect of the volatility bias, we evaluated the EAG responses of female fir coneworm, Dioryctria abietivorella, to five volatile plant compounds, using a new method to normalize EAG responses to account for the loss of antennal sensitivity that occurs over time. Stimuli were generated either from equimolar (uncorrected) solutions or from corrected solutions that were adjusted to yield equimolar airborne concentrations in the air puffed over antennae. When uncorrected solutions were tested, the two most volatile compounds, (E)-2-hexenal and (E)-3-hexenyl acetate, elicited significantly larger EAG responses than three terpenes. When corrected concentrations were tested, the ranking of these responses changed: (E)-2-hexenal elicited significantly smaller EAGs than (−)-α-pinene, (−)-limonene, and (E)-3-hexenyl acetate. On the other hand, there was no effect on the ranking of EAG responses to the two monoterpenes and a sesquiterpene, (−)-trans-caryophyllene, relative to each other. Normalization of EAG data did not affect the overall results (i.e., stimulus rankings) but did reduce their variance within preparations. The results show that when compounds with widely different volatilities are compared in olfactory bioassays, the concentrations of test solutions should be adjusted to produce emissions with equimolar airborne concentrations.

Electroantennogram Responses of a Predator, Perillus bioculatus , and its Prey, Leptinotarsa decemlineata , to Plant Volatiles

The two-spotted stinkbug, Perillus bioculatus, is a predator of the Colorado potato beetle (CPB), Leptinotarsa decemlineata. Behavioral tests revealed that P. bioculatus is attracted to potato plants, Solanum tuberosum L. (Solanaceae), infested by the CPB. Electroantennograms from the antennae of P. bioculatus were recorded in response to compounds present in the headspace of CPB-infested potato plants. (Z)-3-Hexen-1-ol and 2-phenylethanol elicited the highest EAG amplitudes. Linalool, 4,8-dimethyl-1,3(E),7-nonatriene, nonanal, decanal, and (R)-(+)-limonene evoked lower EAG amplitudes. The major headspace components β-caryophyllene and β-selinene produced only weak EAG responses. Antennal sensitivity of the CPB to (Z)-3-hexen-1-ol was higher than that of P. bioculatus, whereas the stinkburg was more sensitive to 2-phenylethanol, β-caryophyllene, (R)-(+)-limonene, and decanal. Among these compounds, 2-phenylethanol is of special interest since it was observed to be emitted by potato foliage only after being damaged by CPBs.

Sex Pheromone of Callosobruchus subinnotatus

Sex pheromone produced by Callosobruchus subinnotatus females stimulates conspecific males to walk upwind toward the source. Gas chromatographic and mass spectrometric analyses of sex pheromone components and their hydrogenated derivatives suggested that the sex pheromone consisted of two short-chain fatty acids, (E)-3-methyl-2-heptenoic acid (E32C7) and (Z)-3-methyl-2-heptenoic acid (Z32C7). The composition of the sex pheromone was confirmed by electrophysiological and behavioral bioassays with synthetic compounds. A mixture of 5 ng E32C7 and 5 ng Z32C7 elicited a 2.1-mV male EAG response, whereas the solvent control elicited 0.1 mV. Mixtures of the two compounds at various ratios were attractive to males in Y-tube bioassays. A pitfall-type trap equipped with a lure impregnated with 1 ng E32C7 and 1 ng Z32C7 was effective in trapping males. Traps baited with the lure caught 80% of males in a glass aquarium during nighttime and 50% during daytime, whereas control traps (solvent treated) in a separate aquarium caught 20% and 10%, respectively.

The EAG Response Spectra of Workers and Drones to Queen Honeybee Mandibular Gland Components: The Evolution of a Social Signal

The EAG Response Spectra of Workers and Drones to Queen Honeybee Mandibular Gland Components: The Evolution of a Social Signal