Dufour Gland Contents of Ants of the Cataglyphis bicolor Group

The species of desert-dwelling ants of the Cataglyphis bicolor (Hymenoptera: Formicidae) group are difficult to distinguish by morphological features. Analysis of the secretion from the Dufour glands of workers of a number of colonies was undertaken to see if it provided a clear test of species. Linked 6c-ms showed in all samples straight and branched-chain alkanes, linear alkenes, ketones, aldehydes, acetates, and a group of C₂₂ to C₂₈ esters not previously identified in this genus. Contents of the Dufour glands of C. savignyi from Tunisia and Egypt were similar, and comprised straight and branched-chain alkanes, alkenes and small amounts of esters. C. bicolor from Tunisia contained compounds similar to C. savignyi but was distinguished from the latter by larger amounts of the esters. The major compound in the glands of C. viaticus was tridecane, in contrast to the pentadecane of other species. It also contained a branched alkane, 3-methyltridecane as a major component. Branched-chain esters and a wide variety of acetates were also found in this species. C. diehlii had a limited range of compounds, with branched alkanes almost completely absent and high proportions of pentadecene and dodecyl acetate. C. bombycinus, a sympatric species, but recognized as not belonging to the bicolor group by its different mandibular gland substances, was notable in having butanoate esters in its Dufour glands. Despite these differences among species, both the great variability of individuals from a single colony and the among between conspecific colonies make species diagnosis from a few individuals difficult, in contrast with postpharyngeal glands, which, as recently reported, give a clearer indication of species.     

Sex pheromone of fall armyworm,Spodoptera frugiperda (J.E. Smith)

Analyses of extracts of pheromone glands and of volatiles from calling female fall armyworm moths,Spodoptera frugiperda (J.E. Smith), revealed the presence of the following compounds: dodecan-1-ol acetate, (Z)-7-dodecen-1-ol acetate, 11-dodecen-1-ol acetate, (Z)-9-tetradecenal, (Z)-9-tetradecen-1-ol acetate, (Z)-11-hexadecenal, and (Z)-11-hexadecen-1-ol acetate. The volatiles emitted by calling females differed from the gland extract in that the two aldehydes were absent. Field tests were conducted with sticky traps baited with rubber septa formulated to release blends with the same component ratios as those emitted by calling females. These tests demonstrated that both (Z)-7-dodecen-1-ol acetate and (Z)-9-tetradecen-1-ol acetate are required for optimum activity and that this blend is a significantly better lure than either virgin females or 25 mg of (Z)-9-dodecen-1-ol acetate in a polyethylene vial, the previously used standard. Addition of the other three acetates found in the volatiles did not significantly increase the effectiveness of the two-component blend as a bait for Pherocon 1C or International Pheromones moth traps.Mention of a commercial or proprietary product does not constitute an endorsement by the USDA.     

Dufour gland contents of three species ofMyrmecia (Hymenoptera: Formicidae), primitive ants of australia

Chemical investigation of the Dufour gland contents of three species ofMyrmecia by GC and GC-MS showed thatM. gulosa andM. nigriceps contain predominantly linear hydrocarbons, whereasM. pilosula contains acetates, aldehydes, and propionates. The major components ofM. gulosa andM. nigriceps are (Z)-8-heptadecene, pentadecane, and heptadecane. Their chemical profiles are similar to each other and to that ofNothomyrmecia macrops, the most primitive living ant, placed by itself in a separate subfamily. The major components ofM. pilosula are hexadecyl acetate, (Z)-9-octadecenyl acetate, (Z)-11-eicosenyl acetate, and (Z)-11-eicosenal. Some geographical variation was observed in the relative proportions of the components. The chemotaxonomic significance for the primitive Australian ant subfamilies Myrmeciinae and Nothomyrmeciinae is considered. The possibility is discussed that 11-eicosenal or geranylcitronellol, minor components of the Dufour gland secretion ofM. gulosa, may be responsible for the aggressive behavior released by the natural stinging secretions.     

Sex pheromone of the avocado pest,Amorbia cuneana (Walsingham) (Lepidoptera: Tortricidae)

The major volatile components in the extract of the female sex pheromone gland ofAmorbia cuneana consisted of (E,E)- and (E,Z)-10,12-tetradecadien-1-ol acetates. The identification was based on electroantennogram bioassay of gas Chromatographic effluent from sex pheromone gland extract, relative retention times on polar and nonpolar gas chromatographic columns, chemical degradation (ozonolysis, saponification), mass spectrometry, chemical synthetic methods, and field tests. Based on mass spectrometry and retention times by capillary gas chromatography, traces of (E)-10-tetradecen-1-ol acetate and 1-tetradecanol acetate were also present in the extract. Traps baited with a combination of synthetic (E,E)- and (E,Z)-10,12-tetradecadien-1-ol acetates caught more males than did traps baited with females.This paper reports the results of research only. Mention of a commercial product in this paper does not constitute a recommendation by the U.S. Department of Agriculture.     

Secretions of Stingless Bees: The Dufour Gland of Nannotrigona testaceicornis

The Dufour gland of Nannotrigona testaceicornis is a large, wide, pear-shaped sac. The gland secretion consists chiefly of the diterpene ester all-trans-geranylgeranyl acetate (64% of the total), together with a complex mixture of small amounts of cyclic ketals; mono-, sesqui-, and diterpene compounds; acetates; and other oxygenated compounds. Samples of N. testaceicornis collected at two sites in Brazil and one in México shared the same composition of their glands, suggesting that the species is uniform over this wide geographical area.     

Pheromones of milkweed bugs (Heteroptera: Lygaeidae) attract wayward plant bugs: Phytocoris mirid sex pheromone

The synthetic aggregation pheromone of the large milkweed bug, Oncopeltus fasciatus (Dallas) (Lygaeinae), also attracted males of the plant bug, Phytocoris difficilis Knight (Miridae). Field testing partial blends against the six-component blend comprising the Oncopeltus pheromone showed that cross-attraction of P. difficilis males was due to synergism between (E)-2-octenyl acetate and (E,E)-2,4-hexadienyl acetate. Hexyl acetate was abundant in the metathoracic scent gland (MSG) secretion of P. difficilis males, but because female P. difficilis could not initially be found in the field, further combinatorial tests were guided by prior research on the pheromones of two Phytocoris species in the western United States. The combination of hexyl, (E)-2-hexenyl, and (E)-2-octenyl acetates was as attractive to P. difficilis males as the milkweed bug pheromone, yet no milkweed bugs were drawn to this blend. Gas chromatographic (GC)-electroantennographic detection (EAD) and GC-mass spectrometric (MS) analyses of female P. difficilis MSGs determined that their secretion contained predominantly hexyl, (E)-2-hexenyl, and (E)-2-octenyl acetates (all strongly EAD-active)—the latter two compounds found only in trace amounts from males—plus five minor female-specific compounds, three of which were EAD-active. (E,E)-2,4-Hexadienyl acetate was not detected from P. difficilis females or males. The blend of the three major components, hexyl, (E)-2-hexenyl, and (E)-2-octenyl acetates (2:1.5:1 by volume), was as attractive as the blend of all six EAD-active compounds identified from females, indicating that this ternary blend constitutes the sex pheromone of P. difficilis. Hexyl acetate with (E)-2-octenyl acetate also attracted males of another species, P. breviusculus Reuter, but addition of (E)-2-hexenyl acetate and/or (E,E)-2,4-hexadienyl acetate inhibited attraction of P. breviusculus males. Attraction of P. difficilis males occurred mainly during the first half of scotophase. The possible neurophysiological basis for this asymmetrical cross-attraction is discussed.     

Volatile secretions of old world army antAenictus rotundatus and chemotaxonomic implications of army ant dufour gland chemistry

The Dufour glands ofAenictus rotundatus contain a complex mixture of terpenoids with geranylgeraniol comprising over 50% of the secretion. Some novel compounds have been tentatively identified as higher homologs of 1,3,3-trimethyl-2,7-dioxabicyclo[2,2,1]heptane based on GC-MS data. The Dufour gland secretion ofA. rotundatus is more similar in composition to the secretions of members of the subfamily Ecitoninae than to its closer relatives from the tribe Dorylini, a result that further complicates studies on the phylogeny of army ants. The mandibular glands ofA. rotundatus contain a mixture of 4-methyl-3-heptanone and limonene in trace amounts, and the well-developed postpygidial glands contain methyl anthranilate only.     

Identification of volatile sex pheromone components released by the southern armyworm,Spodoptera eridania (Cramer)

Analysis of sex pheromone gland extracts and volatile pheromone components collected from the calling female southern armyworm,Spodoptera eridania (Cramer), by high-resolution capillary gas chromatography and mass spectroscopy indicated that a number of 14-carbon mono- and diunsaturated acetates and a monounsaturated 16-carbon acetate were produced. Gland extracts also indicated the presence of (Z)-9-tetradecen-1-ol. However, this compound was not found in collections of volatiles. Field trapping studies indicated that the volatile blend composed of (Z)-9-tetradecen-1-ol acetate (60%), (Z)-9-(E)-12-tetradecadien-1-ol acetate (17%), (Z)-9-(Z)-12-tetradecadien-1-ol acetate (15%), (Z)-9-(E)-11-tetradecadien-1-ol acetate (5%), and (Z)-11-hexadecen-1-ol acetate (3 %) was an effective trap bait for males of this species. The addition of (Z)-9-tetradecen-1-ol to the acetate blends tested resulted in the capture of beet armyworm,S. exigua (Hubner), males which provides further evidence that the alcohol is a pheromone component of this species.     

Volatile communication substances in Dufour's gland of virgin females and old queens of the antFormica polyctena

Dufour's gland, which is present in worker ants ofFormica polyctena as a tiny gland emptying into the short duct from the poison reservoir, was found to be very enlarged in virgin females. In egg-laying queens the gland lobes were reduced in size to slender tubes. The glandular secretion was analyzed by capillary gas chromatography and mass spectrometry. Seventy-four percent of the gland secretion from virgin females consisted of undecane, but this compound was less than 1% in queens. Decane and nonane, making up 3.5% in virgin females, were not found in queens. About 74% of their secretion consisted of the lower hydrocarbons: tridecane, pentadecane, heptadecane, and a nonadecene with tridecane dominating to 50%. Apart from these characteristic differences, almost the same compounds were present in virgin females and queens. Thirty-three compounds were identified from the secretion of virgin females and 38 from queens. An additional compound present in trace amounts in the secretion from queens has not been identified. All the compounds were hydro-carbons with the exception of dodecanol, present in trace amounts in virgin females. The compounds are homologous series of saturated, aliphatic hydrocarbons from nonane to tricosane and corresponding series of monounsaturated and methyl-branched hydrocarbons. Geranylgeraniol and geranyl-geranyl acetate, present in worker ants together with some other alcohols and acetates, were absent in females.     

The dufour gland and the secretion placed on eggs of two species of social wasps,Liostenogaster flavolineata andParischnogaster jacobsoni (vespidae: Stenogastrinae)

The secretion placed on eggs and fed to larvae and the ant guard placed on the nest stalk ofParischnogaster jacobsoni contain the same hydrocarbons and in approximately the same proportions as is found in the Dufour gland. The secretion on eggs is a mixture of the contents of the Dufour gland and nectar. The emulsifying agent is a palmitic acid salt. Similarly, inLiostenogaster flavolineata, the egg secretion is an emulsion of nectar and Dufour gland secretion, which contains alkoxyethanol emulsifiers, found in nature for the first time.     

Identification of Sex Pheromones of Anadevidia peponis and Macdunnoughia confusa and Field Tests of Their Role in Reproductive Isolation of Closely Related Plusiinae Moths

Anadevidia peponis and Macdunnoughia confusa are defoliators of plants in the family Cucurbitaceae and Compositae, respectively, in Japan. GC-MS analyses of crude pheromone gland extracts treated with or without dimethyl disulfide indicated that females of A. peponis produced six monoene acetates and two monoene alcohols and that M. confusa females produced five monoene acetates. These components include (Z)-7-dodecenyl acetate as a major common constituent and three other acetates as minor common constituents. The minor constituents are quite different in blend composition. In addition, with (Z)-7-dodecenyl acetate, an indispensable component for male attraction is (Z)-5-decenyl acetate for A. peponis and (Z)-9-tetradecenyl acetate is essential for M. confusa. Field tests with synthetic lures showed synergistic effects of some other minor components and male attraction of three additional Plusiinae species, Macdunnoughia purissima, Ctenoplusia albostriata, and Chrysodeixis eriosoma, suggesting their reproductive isolation is based in part on pheromonal communication.     

Sex Pheromones of Thysanoplusia intermixta and T. orichalcea: Identification and Field Tests

Thysanoplusia intermixta, which inhabits the temperate zone in Japan, is a defoliator of plants in the family Compositae, Apiaceae, and others. By GC-MS analysis, (Z)-7-dodecenyl acetate, (Z)-7-dodecen-l-ol, (5E, 7Z)-5,7-dodecadienyl acetate and (5E, 7Z)-5,7-dodecadien-l-ol were identified from a pheromone gland extract of the virgin females in a ratio of 18:3:100:13. A lure baited only with the major diene acetate could attract male moths of T. intermixta in the field. Other minor components had a synergistic effect on the attraction of the diene acetate. The same four components were included in a very different ratio of 100:6:11:1 in the gland extract of Thysanoplusia orichalcea, a closely related subtropical species. The monoene and diene acetates are essential for field attraction, and a 100:11 mixture is an optimum lure for T. orichalcea males. However, this mixture, including the diene acetate as a minor component, captured very few T. intermixta males, suggesting the possibility that these two Thysanoplusia species are reproductively isolated from each other by pheromonal communication.     

Female sex pheromone of oriental tobacco budworm,Helicoverpa assulta (Guenee) (Lepidoptera: Noctuidae): Identification and field testing

Analysis of ovipositor washings from virgin femaleHelicoverpa assulta (Guenée) (Lepidoptere: Noctuidae) from Korea by gas chromatography (GC) linked to electroantennography and GC linked to mass spectrometry resulted in the identification of nine compounds, hexadecanal, (Z)-9-hexadecenal, (Z)-11-hexadecenal, hexadecyl acetate, (Z)-9-hexadecenyl acetate, (Z)-11-hexadecenyl acetate, hexadecan-l-ol, (Z)-9-hexadecen-l-ol, and (Z)-11-hexadecen-1-ol. However, ovipositor washings from females from Thailand contained mainly the 16-carbon aldehydes with very small amounts of (Z)-9-hexadecenyl acetate. Field tests conducted in Korea, China, and Thailand indicated that a binary blend of (Z)-9-hexadecenal and (Z)-11-hexadecenal was sufficient for attraction, although the most attractive ratio of compounds varied with location. In Korea a 201 blend of compounds was the most attractive, while in Thailand a 7.51 blend was most attractive. In China both blends of hexadecenal isomers were equally attractive. Addition of the hexadecenyl acetates to the 201 blend of hexadecenals in the ratio of 13.3 increased the trap catch of maleH. assulta compared to lures containing the aldehydes alone in Korea but reduced trap catch in China. Addition of the hexadecenyl acetates to the 7.51 blend of hexadecenals had no significant effect on trap catch in Thailand or China compared to the aldehydes alone. The addition of the 16-carbon alcohols to the aldehydes had a significantly inhibitory effect in all three countries, suggesting they are not pheromone components. Taken together these results indicate thatH. assulta is polymorphic with at least two populations responding to different sex pheromones.     

Differences in oxidase and esterase activities involved in pheromone biosynthesis in two species ofChoristoneura

The released pheromone and the glandular lipids, labeled with [¹⁴C]acetate, were analyzed fromChoristoneura orae andChoristoneura fumiferana budworm moths by thin-layer chromatography and autoradiography. Radiolabeled lipids in the gland appeared to be identical in the two moths with both insects containing high amounts of 11-tetradecenyl acetates. In contrast, theC. orae moths released primarily labeled acetate ester and alcohol, and the spruce budworm moths (C. fumiferana) labeled aldehyde consistent with the known composition of their respective pheromones. The levels of the enzymes responsible for converting the acetate ester into aldehyde were found to be significantly lower in gland extracts fromC. orae moths than fromC. fumiferana moths. These results implicate an acetate esterase and an alcohol oxidase in controlling the composition of the pheromone blend released from closely related budworm species.     

Isolation and identification of sex pheromone ofSymmetrischema tangolias (Gyen) (Lepidoptera: Gelechiidae)

The sex pheromone of the South American potato tuber mothSymmetrischema tangolias (syn.:Symmetrischema plaesiosema) was identified as a 2:1 mixture of (E,Z)-3,7-tetradecadien-1-ol acetate and (E)-3-tetradecen-1-ol acetate by means of dual-column GC, EAG, GC-EAD, GC-MS, NMR, and wind-tunnel bioassays. (Z)-5-Tetradecen-1-ol acetate and (Z)-7-tetradecen-1-ol acetate were also identified in the pheromone gland extract. MaleS. tangolias were able to detect these acetates (EAG), but their addition to the two-component sex pheromone did not improve attractiveness. Field trials in Cajamarca and Cusco, Peru, showed that traps baited with the synthetic sex pheromone were able to catch large numbers of maleS. tangolias.     

Chemical Defense in the Stink Bug Cosmopepla bimaculata

Adult Cosmopepla bimaculata discharge a volatile secretion from paired ventral metathoracic glands (MTG) when disturbed. Collected volatiles were similar in both sexes and consisted of n-tridecane (67%), (E)-2-decenal (12%), (E)-2-decenyl acetate (12%), (E)-2-hexenal (3%), hexyl acetate (2%), n-dodecane (2%), a tridecene isomer (1%), and n-undecane, n-tetradecane, and n-pentadecane (all <1%). In addition, undisturbed males produced a novel insect compound, (E)-8-heneicosene, whose function is unknown. The MTG secretion emerges as an enlarging droplet, which is held in place by a cuticular projection and a pleural scent area consisting of specialized rough cuticle surrounding the gland opening. Insects can selectively discharge from either the right or left gland or both glands simultaneously, can control the amount of fluid ejected, and can resorb the ejected secretion droplet back into the gland reservoir. In feeding trials, killdeer (Charadrius vociferous), starlings (Sturnus vulgaris), robins (Turdus migratorius), and anole lizards (Anolis carolinensis) rejected or demonstrated aversion to feeding on the bugs. Furthermore, bugs that lacked the secretion were more susceptible to predation than bugs with secretion, suggesting that the secretion functions in defense against predators.     

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.     

Contents of poison apparatus and their relation to trail-following in the antDaceton armigerum

The poison gland, which is the source of the trail pheromone inDaceton armigerum, contains a mixture of dimethylpyrazine, trimethylpyrazine and 3-ethyl-2,5-dimethylpyrazine in approximately equal proportions, similar to that of the antManica rubida. The cross-activity in trail-following tests with several other species can thus be explained. The Dufour gland ofD. armigerum contains a mixture of 26 identified hydrocarbons, chiefly 9-tricosene, tricosane, and 9-pentacosene. It also contains very small amounts of two sesquiterpene aldehydes, tetramorine-2 and tetramorine-3, already known in other myrmicine Dufour glands. No volatile substances were detected in the pygidial gland.     

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.     

Premating Behavior of Bombus Confusus Males and Analysis of Their Labial Gland Secretion

Premating behavior in the bumblebee Bombus confusus was studied. Visual searching for females is not the only premating strategy of this species, as was believed earlier. Males of B. confusus have a normally developed labial gland and its secretion is used to mark a perch from which they visually search for females. The labial gland secretion contains geranylcitronellol and (Z)-9-octadecenyl acetate as the main components.