Flight Initiation by Male <Emphasis Type="Italic">Rhodnius prolixus</Emphasis> is Promoted by Female Odors

Several triatomine bug species utilize chemical cues for sexual communication. We tested whether female or male Rhodnius prolixus, a vector of Chagas disease, produce volatile chemicals that elicit flight responses from conspecifics, and then isolated the source of the chemical. Males confronted with an airstream containing female odors showed a significantly greater take-off frequency compared to a blank airstream or an airstream with male odors. In contrast, females exhibited similar take-off frequencies to male or female odor as to a clean airstream. Occlusion of female metasternal glands with paraffin wax resulted in a significant decrease in male take-off frequency compared to that of intact females. Additionally, excised female metasternal glands elicited a similar take-off frequency from males as did intact females, both significantly greater than the take-off frequency to clean air. These results show that R. prolixus females release a pheromone from their metasternal glands that causes upwind flight in conspecific males.     

Larval Diet Affects Male Pheromone Blend in a Laboratory Strain of the Medfly, <Emphasis Type="Italic">Ceratitis capitata</Emphasis> (Diptera: Tephritidae)

The Mediterranean fruit fly (medfly) Ceratitis capitata is a polyphagous pest of fruits and crops with a worldwide distribution. Its ability to use different larval hosts may have multiple effects, including impacts on adult reproductive biology. The male sex pheromone, which plays a key role in attracting both other males to lekking arenas and females for mating, is a mixture of chemical compounds including esters, acids, alkanes and terpenes known to differ between laboratory strains and wild-type populations. The relationship between larval diet and adult pheromone composition remains unexplored. Here, we investigated the effect of larval diet, including laboratory media and fresh fruits, on the composition of the male pheromone mixture. Using Headspace Solid Phase Microextraction we collected the pheromone emitted by males reared as larvae on different substrates and found both qualitative and quantitative differences. A number of alkanes appeared to be typical of the pheromone of males reared on wheat bran-based larval medium, and these may be cuticular hydrocarbons involved in chemical communication. We also detected differences in pheromone composition related to adult male age, suggesting that variations in hormonal levels and/or adult diet could also play a role in determining the chemical profile emitted. Our findings highlight the plasticity of dietary responses of C. capitata, which may be important in determining the interactions of this pest with the environment and with conspecifics. These results also have applied relevance to increase the mating competitiveness of mass-reared C. capitata used in Sterile Insect Technique programs.     

Behavioral and Olfactory Responses of Female <Emphasis Type="Italic">Salaria pavo</Emphasis> (Pisces: Blenniidae) to a Putative Multi-component Male Pheromone

The peacock blenny, Salaria pavo (Risso 1810), typically breeds in rocky shores of the Mediterranean and adjacent Atlantic coast. Males defend a territory around a hole or cavity wherein females deposit eggs that the male guards until hatching. A pair of exocrine glands on the anal fin (anal glands) of males produces a putative pheromone involved in attraction of reproductively competent females to the nest. We used behavioral assays to assess species-specific attraction of reproductively competent females to putative male pheromones, including the anal gland pheromone. Additionally, chromatographic fractions of anal glands and male-conditioned water were tested for olfactory potency in females by electro-olfactogram analysis (EOG). In a flow-through tank or fluviarium, reproductive females were attracted to male-conditioned water and to the anal gland macerate from conspecifics but not to those of a closely related heterospecific. In addition, attraction of reproductive females to conspecific anal gland macerate occurred only during their initial upstream movement in the fluviarium; this was an ephemeral response when compared with the response to male-conditioned water that attracted females throughout the entire period of observation (5 min). Reproductive females also were attracted during the entire period of observation to water-conditioned by conspecific males whose anal glands had been removed. However, the attraction was more variable than that to water conditioned by intact males. Moreover, females were not attracted to male (without anal glands) odor during their initial upstream movement in the fluviarium. Finally, non-reproductive females were not attracted to the conspecific anal gland macerate. The EOG responses of females to molecular weight fractions and solid-phase extraction and high-performance liquid chromatography fractions of anal gland macerates and male-conditioned water (with and without anal glands) suggest that the anal glands release hydrophilic odorants that consist mainly of molecules smaller than 500 Da. Furthermore, males released potent odorants that do not originate from the anal glands. We hypothesize that females respond to a multi-component male pheromone to find mates. The putative anal gland pheromone is possibly comprised of hydrophilic odorants, whereas the other component(s), presumably of gonadal origin, may be less water-soluble. Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

The Response to Selection for Broad Male Response to Female Sex Pheromone and its Implications for Divergence in Close-Range Mating Behavior in the European Corn Borer Moth, Ostrinia nubilalis

Coordinated sexual communication systems, seen in many species of moths, are hypothesized to be under strong stabilizing natural selection. Stabilized communication systems should be resistant to change, but there are examples of species/populations that show great diversification. A possible solution is that it is directional sexual selection on variation in male response that drives evolution. We tested a component of this model by asking whether ‘rare’ males (ca. 5 % of all males in a population) of the European corn borer moth (ECB), Ostrinia nubilalis, that respond to the sex pheromones of both ECB and a different Ostrinia species (O. furnacalis, the Asian corn borer, ACB), might play an important role in diversification. We specifically tested, via artificial selection, whether this broad male response has an evolvable genetic component. We increased the frequency of broad male response from 5 to 70 % in 19 generations, showing that broad-responding males could be important for the evolution of novel communication systems in ECB. We did not find a broader range of mating acceptance of broad males by females of the base population, however, suggesting that broad response would be unlikely to increase in frequency without the involvement of other factors. However, we found that ECB selection-line females accepted a broader range of courting males, including those of ACB, than did females of the base population. Thus, a genetic correlation exists between broad, long-range response to female sex pheromone and the breadth of female acceptance of males at close range. These results are discussed in the context of evolution of novel communication systems in Ostrinia.     

Orientational Behaviors and EAG Responses of Male Codling Moth After Exposure to Synthetic Sex Pheromone from Various Dispensers

The effect of brief pheromone exposures on responses of codling moth (Cydia pomonella L.) males was tested by flight-tunnel and electroantennogram (EAG) studies. Males were preexposed to pheromone for up to 3 min as they sat in release cages or for shorter times (a few seconds to several min) upon initiating flights or orienting in plumes. Brief exposures to Isomate-C Plus dispensers nearly eliminated moth orientations to 0.1 mg codlemone [(E,E)-8,10-dodecadien-1-ol] and 0.1 mg three-component (codlemone/12OH/14OH, 100:20:5 ratio) lures 15 min later. However, there was no associated change in EAG responses between preexposed and control moths. Behavioral responses of Isomate-C Plus-exposed males were normal 24 hr following exposure. The reduced sexual responsiveness observed following exposure to Isomate dispensers appeared to be associated with an elevation of response threshold. Brief preexposure to 0.1 mg codlemone and three-component lures also reduced orientational behavior of males 15 min later, but to a lesser degree than when preexposed to Isomate-C Plus dispensers. Male behavior following preexposure to a 0.1 mg codlemone/pear ester [(2E,4Z)-2,4-decadienoate] lure (1:1 ratio) was no different from exposure to codlemone only. Orientational disruption in plots treated with 10 dispensers of Isomate-C Plus per tree was 88.3 and 95.9% for 1.0 and 0.1 mg codlemone lures, respectively. Some males did orient to 0.1 mg codlemone lures so we caution that flight-tunnel experiments on preexposure may overestimate the actual pheromone exposure dosage received by feral moths in treated orchards. Importantly, this work documents that a portion of feral males within a population has the capacity to overcome communicational disruption by high densities of Isomate-C Plus dispensers.     

Comparative Studies of the Ability of Males to Discriminate Between Sexes in Caloglyphus spp.

Female sex pheromones that trigger the mounting behavior of conspecific males have been identified in four species of the genus Caloglyphus—C. polyphyllae, C. rodriguezi, and two unidentified species. Although the sex pheromones are distributed over both males and females, content ratios of female to male vary from 1.4 in Caloglyphus sp. MJ to 3.4 in C. polyphyllae, 6.3 in C. rodriguezi, and 8.4 in Caloglyphus sp. HP. In the former two species the sex pheromones are also detectable in nymphal stages as a major component, while the pheromones are found to be adult specific in the latter two species. Evaluation of the ability of males to recognize conspecific females revealed that males of all three species, except the Caloglyphus sp. MJ, could discriminate between females and males. We conclude that the differences in the amounts of the female sex pheromone function to facilitate in discrimination by males of conspecific females in the three species, while males of Caloglyphus sp. MJ cannot do this because the 1.4-fold difference is insufficient. It is suggested that the sex pheromone of Caloglyphus spp. might have evolved from a common compound, which originally functioned as a male sexual excitant, into the sex-specific compound that now functions to allow males to discriminate females from males.     

1-Methylethyl Octanoate, a New Lepidopteran Sex Pheromone from the Bagworm, Megalophanes viciella

Extracts obtained by rinsing the thorax and extracts from the tergal sex pheromone gland of females of the bagworm moth, Megalophanes viciella, as well as headspace samples from live animals, were analyzed to reveal the chemical structure of the pheromone. As a result, 1-methylethyl octanoate was established as a main sex pheromone component of this species. This was further confirmed by numerous catches of conspecific males in traps baited with this compound in Bulgaria and Roumania.     

Attraction of male beetles to grubs: Evidence for evolution of a sex pheromone from larval odor

Females of the scarabaeid beetleCyclocephala lurida produce a volatile sex pheromone which attracts conspecific males. Field experiments demonstrated that larvae of both sexes also emit volatile chemicals that stimulate similar responses in adult males, including attempts by the attracted males to mate with the nonreproductive immature stage. Significantly more adult males were caught in traps baited with conspecific male or female larvae or adult females than in blank control traps. Hexane extracts of both male and female grubs were at least as effective as live larvae in trapping male adults, demonstrating that the behavioral responses are mediated by volatile chemicals. Sensory and behavioral responses of males to sex pheromones emitted by adult females are part of the functional communication system. However, their response to grubs is not functional, because grubs are normally temporally and spatially inaccessible to mate-seeking males. In theory, the evolution of a communication system is problematic because it requires the development of a signal in one sex and the sensory and behavioral attributes to respond to that signal in the other sex. The ontogeny of sex pheromone communication inC. lurida suggests a partial solution to this evolutionary problem. We propose that this sex pheromone communication system is probably derived from noncommunicative volatile chemicals that are lost in adult males and retained by adult females.     

Attraction of Female and Male Bactrocera papayae to Conspecific Males Fed with Methyl Eugenol and Attraction of Females to Male Sex Pheromone Components

The attraction of female and male Bactrocera papayae to conspecific males fed with methyl eugenol (ME) and female attraction to male synthetic sex pheromone, trans-coniferyl alcohol (CF), were evaluated in a wind tunnel. Earlier and greater attraction were exhibited by both females and males to ME-fed than to non-ME-fed males as dusk approaches. Males increased their precopulatory behavior (i.e., wing fanning and mounting) during the period of higher attractancy. These data confirm that the consumption of ME enhances the mating competitiveness of males and suggest that ME also functions as a precursor to the male sex and aggregation pheromones. Three phenylpropanoid compounds biosynthesized from ME, coniferyl alcohol, 2-allyl-4,5-dimethoxyphenol, and 3,4-dimethoxycinnamyl alcohol, were detected in male rectal gland along with an endogenous rectal compound, N-(3-methylbutyl) acetamide. When offered singly to the females, coniferyl alcohol was found to be most attractive.     

A Male Sex Pheromone in a Scorpionfly

It has been postulated that males of a number of scorpionfly species produce sex pheromones. This is based on the observation that females often respond only to conspecific males when they evert their genital pouch, the proposed site of pheromone release. In this study, we prove that in Panorpa germanica (Mecoptera, Panorpidae), the eversion of a male’s genital pouch is associated with the release of a volatile sex pheromone. In dual choice situations, females showed a high preference for ‘calling’ (males with everted genital pouch) over noncalling individuals. Volatiles emitted by males and females were collected and identified by coupled gas chromatography–mass spectrometry. Two aldehydes [(2E,6Z)-nona-2,6-dienal and (E)-non-2-enal] were characteristic of calling males but not of noncalling or immature males or females. Bioassays with synthetic compounds confirmed that the identified substances are attractive to females. To the best of our knowledge, this is the first identification of a sex pheromone in scorpionflies.     

Male wing-gland pheromone ofEphestia elutella

Sex pheromone extracted from glands on the forewings of maleEphestia elutella (Hübner) elicits a stereotyped courtship response from conspecific females. A bioassay for this sex pheromone was developed based on this behavior. Maximum production and responsiveness for males and females, respectively, occurred in insects more than 24 hr old.E. elutella females were not responsive to extracts made fromE.figulilella Gregson,E. kuehniella Zeller,E. cautella (Walker), orPlodia interpunctella (Hübner) males.Mention of a commercial or proprietary product does not constitute an endorsement by the USDA.     

Characterization of a Behaviorally Active,Gender-Specific Volatile Compound from the Male Asparagus Fly <Emphasis Type="Italic">Plioreocepta poeciloptera</Emphasis>

Adult male asparagus flies exhibit typical calling behaviors (suggestive of pheromone production) during which they emit a single volatile compound that was identified as isopropyl (S)-5-hydroxyhexanoate. In laboratory bioassays, synthetic samples elicited an arrestant response in females, but did not appear to attract females. On the other hand, the synthetic material attracted conspecific males in olfactometer bioassays.     

Evidence for volatile male-produced pheromone in banana weevilCosmopolites sordidus

Females of the banana weevil,Cosmopolites sordidus, were attracted to and made longer visits to live conspecific males, trapped volatiles from males, and dissected male hindguts in a still-air olfactometer. Male weevils were attracted to volatiles trapped from males and made longer visits to live males and volatiles from males. Live females, collected volatiles from females and female hindguts, elicited small or no behavioral responses from either sex. Electroantennogram (EAG) responses from both male and female antennae were elicited by collected volatiles from males and by dichloromethane extracts of male hindguts and bodies but not by surface washes of males. No significant EAG responses were given to equivalent material from females. It is therefore suggested that male banana weevils release an aggregation pheromone via their hindgut.     

Use of a sex attractant and an inhibitor for monitoring winter moth and bruce spanworm populations

A single sex pherormone component was isolated from abdominal tip extracts of female Bruce spanworm.Operophtera bruceata (Hulst). This was identified as (Z,Z,Z)-1,3,6,9-nonadecatetraene by capillary gas chromatography (GC), electroantennography, and mass spectrometry. In addition, hexane extracts of female abdominal tips from Bruce spanworm and the winter moth.O. brumata L., were analyzed by GC coupled to an electroantennographic detector (GC-EAD). The extracts ofO. bruceata andO. brumata females elicited only a single response, at the same retention time, from antennae of their conspecific and reciprocal males. In field tests conducted in Saskatchewan, traps baited with the synthetic tetraene captured Bruce spanworm males. In tests carried out on Vancouver Island, British Columbia, where the two species coexist, both Bruce spanworm and winter moth males were captured. The attractancy of lures containing the synthetic pheromone alone and in combination with several structurally related analogs was field tested at both locations. One of these, an isomer of the natural pheromone, (E,Z,Z)-1,3,6,9-nonadecatetraene, inhibited the capture of Bruce spanworm males but had no effect upon the number of winter moth males which were taken. Thus, populations of these two very similar species can be distinguished by employing traps baited with pheromone ± the inhibitor. Coneorifice Hara traps were found useful for field trapping males of both species.Issued as NRCC No. 26107     

A new component of the female sex pheromone ofBlattella germanica (L.) (Dictyoptera: Blattellidae) and interaction with other pheromone components

A fourth component, 3,11-dimethyi-2-heptacosanone, was identified as a cuticular contact sex pheromone of the female German cockroach,Blattella germanica. In behavioral assays, higher dosages of 3,11-dimethyl-2-heptacosanone were needed to elicit similar sexual responses in males to those elicited by the major pheromone component, 3,11-dimethyl-2-nonacosanone. A 1585 blend of the C₂₇ and C₂₉ methyl ketone homologs resulted in a dose-response curve intermediate between that of each of the components alone, indicating independence of activity of each component and lack of synergism. Moreover, the activity of 3,11-dimethyl-2-nonacosanone was not enhanced by female cuticular hydrocarbons. The relationship between sexual responses of males to females and to isolated female antennae, and the amount of cuticular pheromone on whole females was investigated. Cuticular sex pheromone found on females increased with the age of the female, as did the male response to whole females. However, a bimodal male response was elicited by isolated female antennae. Differences between behavioral and analytical assays of pheromone are discussed.     

Kairomonal Response of the Parasitoid, <Emphasis Type="Italic">Bracon hebetor</Emphasis> Say,to the Male-Produced Sex Pheromone of Its Host,the Greater Waxmoth, <Emphasis Type="Italic">Galleria mellonella</Emphasis> (L.)

Bracon hebetor is a larval ectoparasitoid that utilizes several pests belonging to the family Pyralidae (Lepidoptera) as hosts. In the present study, we analyzed the kairomonal response of this wasp to the male-produced sex pheromone of a host, the greater wax moth Galleria mellonella, an economically important pest of honeybees, Apis mellifera. Coupled gas chromatography-electroantennographic detection (GC-EAD) revealed three compounds in headspace collections from male G. mellonella that elicited responses from B. hebetor antennae: decanal and the previously identified sex pheromone components, nonanal and undecanal. Y-tube olfactometer tests that used naïve, mated wasps showed that females, but not males, were highly attracted to (a) male G. mellonella headspace samples, (b) two synthetic blends of nonanal and undecanal (in ratios matching that found in male moth samples), and (c) the two aldehydes tested individually. Further, female wasps did not discriminate between a blend of aldehydes and male G. mellonella headspace. In dose-response trials that used octanal, nonanal, decanal, and undecanal, no difference in EAG responses of the two sexes was observed, except for undecanal at the second highest dose, for which female antennae showed significantly larger responses than did male antennae. When the two binary blends were tested at different doses, female wasps were significantly attracted to the two highest doses (1 µg and 10 µg), but not to the lowest dose (100 ng). Our results show that females of this economically important parasitoid utilize the male-produced sex pheromone of a host as an indirect cue to guide them to potential oviposition sites.     

Male bugs modulate pheromone emission in response to vibratory signals from conspecifics

Male Nezara viridula emit a volatile sex pheromone that acts as a long-range attractant to females. Both sexes communicate through vibrations once they are on the same plant. Males respond to the female calling song by emitting a male courtship song, and they orient to the female calling song on a plant. Simultaneity between vibratory and chemical communication during the last steps of mate finding suggests that pheromone emission might be modulated by signals from conspecifics. Male volatile emissions were collected with fibers for solid-phase microextraction, while male bugs were stimulated with natural and artificial signals. Percentages of males releasing pheromone and collected amounts of pheromone increased when males were stimulated with female calling song. Pheromone emission was stable in males stimulated with male rivalry songs, and it decreased in males stimulated with a 100-Hz artificial signal. The ability of male bugs to modulate their pheromone emission may reduce metabolic costs, reduce parasitism, and offer a better synchronization of sexual activity.     

Sex Pheromone of Glossina tachinoides: Isolation, Identification, and Synthesis

Study of lipids from male and female laboratory-reared flies led to the demonstration of a potent contact sex stimulant in extracts and cuticular hydrocarbons of the female tsetse fly Glossina tachinoides (Westwood) against conspecific males. Thin-layer and column chromatography indicated that extracts contained hydrocarbons and saponifiable lipids. Biological activity was found in the alkanes from females, including prominent branched-chain alkanes that were detected by gas chromatography (GC). The alkanes were separated and collected by preparative gas chromatography (GC), and only the 37-carbon region showed biological activity. GC–mass spectrometry showed the major peak contained a mixture of isomeric 11,23-, 13,25- plus a minor amount of 11,21-dimethylheptatriacontane. Two racemic isomers were synthesized, and bioassays showed that the greatest activity was possessed by the 11,23- isomer with somewhat less activity in 13,25-dimethyl heptatria-contane. Dose–response data showed ED₅₀ at 5 g per decoy with solvent-washed males, nonspecific females, or corks as decoys. These alkanes released sexual activity in males that comprised most of the behaviors released by a female fly of the same species.     

Odor Preferences of Weanling and Mature Male and Female Pine Voles

Olfactory cues from conspecifics have strong influences on behavior. Both female and male juveniles are attracted to familiar odors while adults show preferences for odors of opposite-sex conspecifics. Females show an increased preference for male odors as they mature, but no study has examined whether change with age is seen in males. In this experiment, responses of weanling and mature male and female pine voles (Microtus pinetorum) to familiar odor cues from their family versus those of an unfamiliar, opposite-sex conspecific were examined. Voles were tested in a three-chambered cage with end chambers containing the stimuli. There was a significant difference in odor preference between weanling and mature females. Weanling females preferred odor cues from their families, and mature females preferred the odors of unfamiliar mature males. The pattern was similar in males but the effect was not as strong. The results support the hypothesis that responses to odor cues change with age in pine voles. At least for females, the change in response to odor cues may be involved in the timing of reproduction and thus dispersal.     

Identification,Synthesis, and Behavioral Activity of 5,11-Dimethylpentacosane,A Novel Sex Pheromone Component of the Greater Wax Moth,Galleria Mellonella (L.)

The greater wax moth, Galleria mellonella (L.), is a serious and widespread pest of the honeybee, Apis mellifera L. In contrast to most moths, for which long-range mate finding is mediated by female-produced sex pheromones, G. mellonella males attract conspecific females over long distances by emitting large amounts of a characteristic scent in combination with bursts of ultrasonic calls. The male scent for this species was previously identified as a blend of nonanal and undecanal. When these compounds were bioassayed, characteristic short-range sexual behavior, including wing fanning, was triggered in conspecific females, but the aldehyde blend failed to elicit attraction over longer distances. We identified, via analysis and synthesis, a third male-specific compound, 5,11-dimethylpentacosane. We show that it acts as a behavioral synergist to the aldehydes. In wind tunnel experiments, very few female moths responded to the aldehyde blend or to 5,11-dimethylpentacosane tested separately, but consistently showed orientation and source contact when a combination of all three compounds was applied. The level of attraction to the three-component mixture was still lower than that to male extract, indicating that the composition of compounds in the synthetic blend is suboptimal, or that additional pheromone components of G. mellonella are yet to be identified. The identification of 5,11-dimethylpentacosane is an important step for the development of an efficient long-range attractant that will be integrated with other environmentally safe strategies to reduce damage to beehives caused by wax moths.