Chemical composition of North American bee propolis and biological activity towards larvae of greater wax moth (Lepidoptera: Pyralidae)

Bee propolis is a sticky amalgamation of plant resins collected by honeybees (Apis mellifera L.) and used in the hive for filling cracks and repairing combs. Propolis contains a diversity of compounds of plant origin, and is reported to have medicinal, antimicrobial, insecticidal, and phytotoxic properties. We examined the physical and chemical composition of North American samples of bee propolis from several sites in North America and tested for bioactivity against larvae of the greater wax moth (Galleria mellonella L.), a common apiary pest. The amount of methanol-extractable resin in samples from Ohio and Georgia ranged from 24% to 79% by weight. Propolis collected from hives in Ohio was more chemically diverse (over 30 compounds detected by paper chromatography) than material from south Georgia (fewer than 10 major compounds) and contained a lower proportion of methanol-insoluble beeswax. The paper chromatographic surveys revealed little variation in the chemical profile of specific hives over a six-month period and no differences between propolis from adjacent hives. Four flavonoids were identified from propolis collected in Ohio: kaempferol, galangin, 3,3-dimethoxyquercetin and 3-methoxykaempferol. When mixed into artificial diet, fractionated propolis reduced larval growth of the greater wax moth, but not dramatically. An array of phenolics reported from propolis (caffeic acid, chrysin, ferulic acid, galangin, kaempferol, and quercetin) were bioassayed individually for effects on larvae, but none reduced larval growth at the concentrations tested, suggesting that wax moths are tolerant of some phenolics in their diet.     

Sex pheromone of the european sunflower moth,Homoeosoma nebulellum (Den. & Schiff.) (Lepidoptera: Pyralidae)

Four components, (Z)-9-tetradecenal (8.6%), (Z,E)-9,12-tetradecadienal (4.8%), (Z)-11-hexadecenal (49.5%), and (Z)-13-octadecenal (37.1%), were identified in extracts of female pheromone glands of the European sunflower moth,Homoeosoma nebulellum (Lepidoptera: Pyralidae) using GC and GC-MS analyses. EAG and single-cell recordings of male antennal receptors gave strong evidence for (Z,E)-9,12-tetradecadienal as the antennal key compound of sex pheromone detection in this species. This result was confirmed by field trapping; removal of (Z,E)-9,12-tetradecadienal from quaternary blends completely suppressed the male catches. The synthetic blends with this compound as a major component caught five times less males than the blends reproducing the ratio found in the female extracts [5% of (Z,E)-9,12-tetradecadienal only]. The occurrence of a minor component perceived as the most biologically relevant compound is discussed.     

Female sex pheromone components of jasmine mothPalpita unionalis (Lepidoptera: Pyralidae)

(E)-11-Hexadecenal and (E)-11-hexadecen-1-yl acetate were found in abdomen tip extracts from females of the jasmine mothPalpita unionalis (Hübn). The identification was based on capillary GC analyses, mass spectrometry, and laboratory and field tests. In laboratory bioassays, both components elicited a low level of upwind flight by males. The two components when tested separately in the field were inactive, but the blend of the two components at a ratio of (3:7) was highly attractive to males. Traps baited with 1 mg of the two-component blend were competitive to traps baited with five virgin females. The addition of Z isomers components reduced male capture.     

In vitro maintenance of the sex pheromone gland of the female Indian meal mothPlodia interpunctella (Hübner)

Sex pheromone glands ofPlodia interpunctella were isolated from surface-sterilized donors of different ages, freed of most of the attached gut, oviduct, and other tissues; rinsed in sterile medium; and cultured in 1 ml of culture medium. The sex pheromone gland cells that were cultured for 10 days in either chemically defined Grace's medium or modified Grace's medium appeared normal in histological examinations. Bioassays of extracted medium in which pheromone glands from mature females had been incubated showed that a greater percentage of the available pheromone was recovered from modified Grace's medium than from chemically defined Grace's medium.Lepidoptera: Pyralidae.This research was supported in part by NIH Grant 1-F34 GM 06251.     

Sex pheromone of cranberry fruitworm,Acrobasis vaccinii riley (Lepidoptera: Pyralidae)

The following compounds and (approximate ratios) were identified in sex pheromone gland extracts of femaleAcrobasis vaccinii Riley by comparison of gas chromatography-mass spectrometric traces with those of synthetic standards: (E,Z)-, (Z,E)-, (Z,Z), and (E,E)-8, 10-pentadecadien-l-ol acetates (100:1:2:12), a dodecen-l-ol acetate (8), (Z)-8-, (Z)-9-, and (E)-9-pentadecen-l-ol acetates (3:23:4), two heptadecen-l-ol acetates (4:4), tetradecyl, pentadecyl, hexadecyl, and heptadecyl acetates (3:15:10:8), dodecan-l-ol (6), tetradecan-l-ol (5), and hexadecan-l-ol (23). The amount of (E,Z)-8, 10-pentadecadien-l-ol acetate (E8,Z10–15:Ac) in the extract was about 0.5 ng/female. Electroantennographic analysis of gas chromatographic fractions of female sex pheromone gland extract showed that the fraction containingE8,Z10–15:Ac elicited the greatest response. Alone,E8,Z10–15:Ac failed to elicit upwind flight of males in flight-tunnel tests, and traps baited with it did not catch males in field experiments. WhenE8,Z10–15:Ac was combined with (E)-9-pentadecen-l-ol acetate (100:4), male upwind flight response in flight-tunnel tests was equivalent to those obtained with extract of female sex pheromone glands (synthetic, 62%; natural, 51%), but the percent of males flying upwind that contacted the source was lower (synthetic, 47%; natural, 88%). The lower percent of source contact elicited by the synthetic pheromone could be a result of the difference in isomer ratios of 8,10–15:Ac in the natural and synthetic pheromone or could indicate that the synthetic pheromone is incomplete. Traps baited with the 100:4 combination caught large numbers of males in field experiments.     

Sex pheromone components from asian corn borer,Ostrinia furnacalis (Hubner) (Lepidoptera: Pyralidae) in Taiwan

The female sex pheromone of the Asian com borer,Ostrinia furnacalis, widespread in Taiwan, was confirmed as (Z)-12-tetradecenyl acetate and its geometric isomer (E)-12-tetradecenyl acetate in a ratio of ca. 31 by gas chromatography and gas chromatography-mass spectrometry in selected ion monitoring mode. Males were attracted by the mixture of these two synthetic components in the field, but the attractiveness was less than by virgin females. The presence of minor components in the sex pheromone was therefore suggested.     

Constituents of wing gland and abdominal hair pencil secretions of male African sugarcane borer,Eldana saccharina walker (Lepidoptera: Pyralidae)

In addition totrans-3,7-dimethyl-6-octen-4-olide (eldanolide), vanillin, and 4-hydroxybenzaldehyde, identified by French workers in the wing gland and abdominal hair pencil secretions of the male African sugarcane borer,Eldana saccharina, we have, in an earlier note, reported the presence of several other terpenoid, aromatic, and unbranched-chain compounds such as, (Z)-3,7-dimethylocta-2,6-dienoic acid, 6,10,14-trimethyl-2-pentadecanol, 4-hydroxy-3-methoxybenzyl alcohol, 1-octadecane thiol, 16-hexadecanolide, and 18-octadecanolide in these secretions. In the present paper experimental details and spectral evidence supporting the identification of these compounds, as well as the identification of (Z)-9-hexadecenal and cw-3,7-di-methyl-6-octen-4-olide (cis-eldanolide), are reported. Using electroantennography it was found that male and female antennae reacted approximately equally strongly to both secretions. This result was confirmed in analyses of the secretions using coupled gas chromatography-electroantennography and it was found that male as well as female antennae responded to eldanolide. Vanillin, substituted phenols related to vanillin, and some oxygenated monoterpenes elicited weak responses in male and female antennae. In some analyses 6,10,14-trimethyl-2-pentadecanol, present in the secretions of the insect, gave a strong antennal response. The results obtained in dynamic and static headspace determinations showed that several of the organic compounds present in the glandular secretions are released in detectable quantities and are present in widely varying quantitative ratios in the effluvia of individual calling male moths.     

Development of pheromone lure for monitoring field populations ofEoreuma loftini (Lepidoptera: Pyralidae)

Formulations of Mexican rice borer (MRB) pheromone components were more effective in capturing males in fields of sugarcane when dispensed from rubber septa than when evaporated from four other substrates. Concentrations of 0.63–10.0 mg of total pheromone per lure were effective over a 112-day period when formulated in rubber septa in a ratio of 811.3 of (Z)-13-octadecenyl acetate, (Z)-11-hexadecenyl acetate, and (Z)-13-octadecenal, respectively. Component ratios of 411.3 and 811.3 were equally effective in capturing male MRB, but efficacy decreased with higher relative concentrations of (Z)-13-octadecenyl acetate.Mention of a commercial or proprietary product does not constitute an endorsement by the USDA.     

Identification of sex pheromone of browntail moth,Euproctis chrysorrhoea (L.) (Lepidoptera: Lymantriidae)

A unique sex attractant pheromone was isolated and identified from extracts of ovipositor tips from the female browntail moth (Lepidoptera: Lymantriidae). The pheromone compound, (Z,Z,Z,Z)-7,13,16,19-docosatetraen-1-ol isobutyrate, CH₃CH₂CH=CHCH₂CH=CHCH₂CH= CH(CH₂)₄CH=CH(CH₂)₆O₂CCH(CH₃)₂, was identified by a combination of gas chromatography, mass spectrometry, and microreactions and was confirmed by synthesis. Traps baited with 5–50 g of the synthetic pheromone, dispensed from rubber septa treated with an antioxidant and a UV stabilizer, gave male moth captures that were comparable to traps baited with three virgin females. Higher (250 g) and lower (0.04–2.5 g quantities of the synthetic pheromone on septa captured somewhat fewer males. Captures were the same for similar quantities of the natural and synthetic pheromone. Minor, inactive components in the tip extract were identified as a mixture of 7- and 8-pentacosanone.     

Female sex pheromone of the pickleworm,Diaphania nitidalis (Lepidoptera: Pyralidae)

Heptane extracts of the ovipositors from pickleworm adults (Diaphania nitidalis) were found to contain (E)-11-hexadecenal along with proportionally smaller amounts of (Z)-11-hexadecenal, (E)- and (Z)-11-hexadecen-1-ol, hexadecanol, hexadecanal, and a trace amount of (E,Z)-10,12-hexadecadienal. Assays conducted in a flight tunnel and in the field showed that a synthetic mixture of the five unsaturated compounds elicited behavioral responses from pickleworm males that were indistinguishable from those elicited by extracts of the female or by mate-calling females. When any component was deleted from the set of five unsaturated compounds, the intensity and extent of male responses to the resulting mixtures were significantly attenuated. The female sex pheromone of the pickleworm resembles the pheromone of a congeneric species,D. hyalinata, but bioassays indicated that (E,E)-10,12-hexadecadienal, produced byD. hyalinata but not by the pickleworm, plays a role in pheromonal specificity.     

Sex pheromone of female african white rice stem borer,Maliarpha separatella (Lepidoptera: Pyralidae) from Sierra Leone: Identification and field testing

Analysis of ovipositor washings of femaleMaliarpha separatella from Sierra Leone by high-resolution gas chromatography (GC) linked to a male electroantennograph (EAG) indicated the presence of three electrophysiologically active compounds. The GC retention times of these compounds were consistent with those of (Z)-9-tetradecen-1-ol, (Z,E)-9,12-tetradecadien-1-ol, and (E,E)-10,12-tetradecadien-1-ol. Analysis by El and CI mass spectrometry of ovipositor washings confirmed these identifications and also indicated the presence of the saturated analog, tetradecan-1-ol. There was no evidence, from these analyses, of the corresponding aldehydes or acetates. The EAG-active compounds were present in the ovipositor washings in a ratio of approximately 23.51, with the major component constituting approximately 0.4 ng per ovipositor. GC-MS analysis of entrained female effluvia confirmed that the EAG-active compounds were released by virgin females. Field testing of the EAG-active compounds indicated that (Z,E)-9,12-tetradecadien-1-ol and (E,E)10,12-tetradecadien-1-ol were attractive to male moths in ratios of between 9:1 and 39:1, while (Z)-9-tetradecen-1-ol was found to reduce trap catch when added to blends of the other two compounds.     

Anemotactic response threshold of the Indian meal moth,Plodia interpunctella (Hübner) (Lepidoptera: Pyralidae), to its sex pheromone

The effect of different concentrations of the sex pheromone (Z,E)-9,12-tetradecadien-1-ol acetate on the upwind anemotactic behavior of the malePlodia interpunctella (Hübner) was measured at 23 ± 1 ° C and 34 ± 1 ° C. The stimulus-response regression lines were analyzed by a new procedure that accounts both for control responses in the absence of pheromone and also for peak responses below 100% in the presence of concentrations considerably above the normal physiological levels. From the regression line for each temperature, the upwind anemotactic thresholds were calculated to be 1.34 × 10⁶ molecules/cm³ at 23 ° C and 1.65 × 10⁴ molecules/cm³ at 34 ° C, similar to other thresholds reported in the literature. Since departures from the two lines occurred at the highest concentrations tested, near 10⁸ molecules/cm³, the upwind anemotactic behavior may change qualitatively above an altered-behavior threshold that is about two orders of magnitude higher than the upwind anemotactic threshold. The lower response at 23 ° C suggests that cool temperatures inhibit flight in response to pheromonal stimulation.Mention of a commercial or proprietary product in this paper does not constitute an endorsement of that product by the USDA.     

Sex pheromone of the cranberry girdler,Chrysoteuchia topiaria (Zeller) (Lepidoptera: Pyralidae)

Z-11-Hexadecen-1-al (Z11-16A1), free of theE isomer, was identified in extracts of femaleChrysoteuchia topiaria (Zeller) abdominal tips. In commercial grass seed fields, traps baited with synthetic Z11-16A1 were almost as attractive as female-baited traps. An alcohol, thought to be Z-11-hexadecen-1-ol on the basis of gas Chromatographic retention times, was also isolated from the extract.     

Female sex pheromone of the melonworm,Diaphania hyalinata (Lepidoptera: Pyralidae), and analysis of male responses to pheromone in a flight tunnel

Ten C₁₆ chain-length compounds were identified from heptane extracts of ovipositors of female melonworm,Diaphania hyalinata (L.). The major constituents of the extracts were (E)-11-hexadecenal and (E,E)-10,12-hexadecadienal [(E,E)-10,12–16:Ald] and the alcohols and acetates of these olefins were found in trace amounts (<2%). Extracts also contained traces of (E,Z)- and (Z,Z)-10,12-16:Ald, hexadecanal, and 1-hexadecanol. Analysis of the behavioral responses of males to synthetic mixtures of these compounds and responses to ovipositor extracts in a flight tunnel showed that a synthetic mixture of the 10 compounds elicited a behavioral repertoire from males that was indistinguishable from that elicited by ovipositor extract. Flight tunnel studies also indicated that six of the 10 compounds probably represent the essential components of the female's sex pheromone.Supported in part by USDA Cooperative Agreement No. 58-32U4-1-299. Scientific Article No. A-4073, Contribution No. 7058 of the Maryland Agricultural Experiment Station. Mention of a commercial product in this paper does not constitute a recommendation by the U.S. Department of Agriculture.     

Variation in relative quantities of airborne sex pheromone components from individual femaleEphestia cautella (Lepidoptera: Pyralidae)

The airborne sex pheromone components (Z,E)-9,12-tetradeca-dien-1-yl acetate and (Z)-9-tetradecen-1-y1 acetate from single calling females ofEphestia cautella (Walker) were trapped within glass capillary tubes and were measured by gas chromatography-mass spectrometry. Broad and similar distributions of relative quantities were found for a laboratory strain and three Australian field strains, and means differed strongly from those reported previously for this species. The overall mean proportion of the two components found for Australian females was 8812. The composition in individuals ranged from 6327 to 973. The proportions for individuals appeared to vary slightly in a random fashion from day to day, and proportions for first-generation progeny were influenced by the maternal blend.Author order in this paper is alphabetical     

Sex pheromone of femaleMyelois cribrella Hübner (Lepidoptera: Pyralidae)

Females ofMyelois cribrella contain about 20 ng/gland of the primary sex pheromone components (Z)-9-tetradecenyl acetate, (9Z,12E)-9,12-tetradecadienyl acetate, and (Z)-11-hexadecenyl acetate in proportions of 4115, respectively. These physiologically active components are accompanied by a number of related compounds such as (Z)-9-tetradecen-1-ol, hexadecyl acetate, (9Z,12E)-9,12-tetradecadien-1-ol, (Z)-11-hexadecen-1-ol, octadecyl acetate, octadecan-1-ol, and eicosyl and docosyl acetates. Octadecyl acetate, the most abundant component, represents about 42 ng/female moth; however, no physiological activity could be attributed to it. In field tests, a trap baited with a 1-mg mixture of (Z)-9-tetradecenyl acetate, (9Z,12E)-9,12-tetradecadienyl acetate, and (Z)-11-hexadecenyl acetate in a ratio of 121 caught more male moths than three live female moths.Pheromones, 58. Pheromones, 57: Bestmann et al. (1987).     

Postembryonic development and reproduction inCorcyra cephalonica (Stainton) (Lepidoptera: Pyralidae) on exposure to eucalyptus and neem oil volatiles

Postembryonic development and adult emergence ofCorcyra cephalonica (Stainton) were adversely affected in varying degrees when individuals were reared for the first 2 weeks of larval life or for a similar duration from the sixteenth day of their lives in an environment of eucalyptus oil volatiles. This was, however, not so in the progeny of parents exposed to these volatiles for only 5 min. Exposure of this pest to neem oil volatiles during larval stages for similar periods failed to produce such adverse effects. A marked decline in the reproductive potential, in terms of egg output and egg hatchability, of the moth was observed when the larvae were reared for the first 15 days in the presence of eucalyptus oil volatiles or when the parents were exposed for 5 min to such an environment during adult life. Thus, a carry over of the detrimental effect of the volatile(s) of this oil on the reproductive potential of the pyralid was indicated.     

Sex pheromone of the pea moth,Cydia nigricana (F.) (Lepidoptera: Olethreutidae)

The sex pheromone of the pea moth,Cydia nigricana (F.), was identified as (E,E)-8,10-dodecadien-1-ylacetate (E,E8,10–12Ac) (approximately 0.1 ng/abdominal tip), in vacuum distillates of virgin female abdominal tips and volatiles emitted by calling females, from its chemical properties and capillary gas chromatography and mass spectrometry data. SyntheticE,E8,10–12Ac and the natural pheromone elicited similar quantitative electrophysiological and behavioral responses from male moths. Other related compounds which also attract male moths in the field,viz., (E)-10-dodecen-1-yl acetate, (E,E)-8,10-dodecadien-1-ol, and (E,E)-8,10-dodecadienal, were not detected in the pheromone andE,E8,10–12Ac was not found in male moths.     

Squash leaf glandular trichome volatiles: Identification and influence on behavior of female pickleworm moth [Diaphania nitidalis (Stoll.)] (Lepidoptera: Pyralidae)

Fourteen volatile compounds occurring in leaf trichomes of yellow squash (Cucurbita pepo L. cv. Early Prolific Straightneck) were identified. These compounds accounted for 83.5% of the volatile matrix. Ubiquitous constituents of the epidermis (myristic, palmitic, and stearic acids,n-tricosane, andn-pentacosane) accounted for 73.7%; these compounds were not bioassayed. The volatileso-,m-, andp-xylene, toluene, 2-heptanone, (R)-(+)- and (S)-(–)-limonene, and germacrene D were tested for their influence on attraction and oviposition by the pickleworm moth (Diaphania nitidalis Stoll.). No single compound, except germacrene D, was attractive. (R)-(+)-Limonene and 2-heptanone were weakly repellent. Mixtures of the highly volatile fractions were as attractive as volatiles emanating from whole, intact leaves. Oviposition levels on treated artificial sites corresponded with levels of visitation. Oviposition was significantly stimulated by whole-leaf volatiles, and (S)-(–)-limonene caused a slight but significant reduction.Mention of a trade name or proprietary product does not constitute a warranty or guarantee by the U.S. Department of Agriculture, nor does it imply exclusion of other products that may also be suitable.     

Identification of sex pheromone components from pheromone gland volatiles of the tomato looper,Plusia chalcites (Esp.)

The sex pheromone glands ofPlusia chalcites release, dodecyl acetate, (Z)-7-dodecenyl acetate, (Z)-9-dodecenyl acetate, 11-dodecenyl acetate, (Z)-8-tridecenyl acetate, and (Z)-9-tetradecenyl acetate. A combination of capillary GC, GC-MS, and dimethyl disulfide derivatization enabled a rigorous identification of all these compounds, some of which were previously found in gland extracts. Bioassays in a flight tunnel showed that a ternary blend of (Z)-7-dodecenyl acetate, (Z)-9-dodecenyl acetate, and (Z)-9-tetradecenyl acetate elicited directed flights from 85 to 100% of the males tested and elicited copulation attempts, at the end of the flights, from 44 to 74% of the males tested. This blend was equal in activity to the natural gland extract. Addition of the other acetates had only a slight influence on the activity of this mixture. Substitution of either (Z)-9-dodecenyl acetate or (Z)-9-tetradecenyl acetate in this blend by 11-dodecenyl acetate gave two ternary mixtures which also elicited high levels of courtship activity, almost as high as that of the original blend. Addition of (Z)-7-dodecenyl alcohol inhibited almost totally the flight activity of males.Lepidoptera: Noctuidae.Contribution from the Agricultural Research Organization (ARO) No. 1701-E, 1986 series.