Surface lipids of social waspPolistes melricus say and its nest and nest pedicel and their relation to nestmate recognition

In eight replicate laboratory tests wherePolistes metricus adults were allowed to choose between their own nest, a second nest, and neither nest, they selected their own nest 66% of the observed time. When the surface hydrocarbons had been extracted from the nests, the wasps chose their own nest only 8% of the time, but after the hydrocarbons were reapplied to the surface of the respective nests, they selected their own nest 47% of the time. These changes are significant. The cuticular lipids were analyzed from individualP. metricus adult females collected from 13 colonies. Surface lipids were recovered from the paper and pedicels of their nests. Eighteen hydrocarbons were identified in these lipid fractions. The major components of the wasp cuticular lipids weren-heptacosane,n-nonacosane, methylhentriacontane, and methyltritriacontane. Factor analysis revealed that extracts of pedicels are all similar in composition, while cuticle and paper extracts vary, sometimes similarly according to colony identity.     

Surface lipids of the social waspPolistes annularis (L.) and its nest and nest pedicel

Cuticular lipids were recovered fromPolistes annularis workers and characterized by combined gas-chromatography-mass spectrometry. These lipids were found to be straight-chain and methyl-branched hydrocarbons with the major components being 13,17-dimethylhentriacontane (18%), 3-methyl-nonacosane (13%), and 3-methylheptacosane (11%). Cuticular lipids with a very similar composition were found on the surface of the larvae, eggs, and adult males ofPolistes annularis as well as on the surface of the nest and nest pedicel of the wasp. Hexadecanoic acid and octadecenoic acid were also found on the surface of the nest pedicel and these acids and/or the hydrocarbons may play a defensive role for the wasps.     

Cuticular lipid profiles of queens,workers, and males of social waspPolistes metricus say are colony-specific

The cuticular lipids ofPolistes metricus queens, workers and males from seven laboratory-maintained colonies were extracted and analyzed by combined gas chromatography-mass spectrometry. Males had higher proportions of alkenes (20.5%) in their cuticular lipids than did queens (2.3%) or workers (7.7%). Discriminant analyses of the cuticular lipid profiles of the adult wasps showed that males group separately from females. Additional analyses showed that queens group with their respective workers by colony and that queens group even more closely with males by colony. The most distinct groupings occurred with workers only by colony and with males only by colony. Stepwise discriminant analyses showed that each type of grouping was dependent upon a different combination of cuticular lipids.     

Cuticular hydrocarbons of the paper wasp,Polistes fuscatus: A search for recognition pheromones

The cuticular chemicals of 124 individual wasps (foundresses and workers) from 23 colonies ofPolistes fuscatus were analyzed. The compounds identified, all of which were hydrocarbons, were similar to those of other vespid wasps in that the bulk of the hydrocarbons were 23–33 carbons in chain length. However, the hydrocarbon profile ofP. fuscatus differed from those of its congeners in its proportions of straight-chain alkanes, methylalkanes, and alkenes. Three of the 20 identified hydrocarbons, 13- and 15-MeC₃₁, 11,15- and 13,17-diMeC₃₁, and 13-, 15-, and 17-MeC₃₃, had properties postulated for recognition pheromones: colony specificity, efficacy in assigning wasps to the appropriate colony, heritability, lack of differences between foundresses and workers, and distinctive stereochemistry.     

Ultrastructural and chemical examination of paper and pedicel from laboratory and field nests of the social waspPolistes metricus say

The ultrastructure and chemical composition of paper and pedicel from nests ofPolistes metricus that were constructed in the laboratory from known building material were compared to paper and pedicel from nests constructed in the field. Scanning electron micrographs showed the addition of a gluelike secretion from the wasp to the construction material. Solid-state [¹³C]NMR, elemental analyses, and amino acid analyses indicate that this secretion is a silklike protein with serine, glycine, alanine, and proline comprising 65–73% of the identified residues.     

Sex attraction in paper wasp,Polistes exclamans viereck (Hymenoptera: Vespidae), in a wind tunnel

A wind-tunnel bioassay was developed to test for the presence of sex attractants in the paper wasp,Polistes exclamans Viereck (Hymenoptera: Vespidae). Males exhibited significant upwind flight and attraction (chemoanemotaxis) in response to airflow passed over unmated females, and to hexane extracts of whole bodies and thoraces of unmated females. Unmated females were attracted to hexane extracts of males and to hexane extracts of each body tagma of males, suggesting distribution of the pheromone over the cuticle by grooming. The ectal mandibular and seventh sternal glands are the likely sources of the male-produced sex attractant(s) since extracts of each elicited female attraction in the flight tunnel. These glands are associated with gastral and mandibular rubbing of perch sites by territorial males ofPolistes species.     

Characterization of pedicel,paper, and larval silk from nest ofPolistes annularis (L.)

The pedicel, nest paper, and larval silk ofPolistes annularis nests were analyzed by high-resolution solid-state [¹³C]NMR. The pedicel was found to have a high nitrogen content (11%), and the NMR spectra indicated that it is a mixture of carbohydrate and protein. The pedicel protein has an amino acid composition that is very rich in glycine, alanine, serine, and proline (67% of identified residues), similar to that of some insect silks. Solid-state [¹³C]NMR indicated that the nest paper is composed predominantly of cellulose. Silk, spun by matureP. annularis larvae, was shown by [¹³C] NMR and amino acid analysis to be a protein very high in serine and alanine (53%), but the amino acid composition is distinct from that of the pedicel protein.     

Alarm response to venom by social waspsPolistes exclamans andP. fuscatus (Hymenoptera: Vespidae)

The venoms ofPolistes exclamans andP. fuscatus elicit alarm behavior and attract attacking wasps. The response is not species specific, for both hetero- and conspecific venoms elicit similar responses in both species. A test in a wind tunnel provided no support for the hypothesis that alarmed wasps release an alarm pheromone on the nest.     

Temporal changes in colony cuticular hydrocarbon patterns ofSolenopsis invicta

Heritable cuticular hydrocarbon patterns ofSolenopsis invicta workers are consistent within colonies for a given sampling time but vary sufficiently from colony to colony to distinguish the colonies from each other. In addition, cuticular hydrocarbon patterns change within colonies over time. Nestmate recognition cues found on the individual's cuticle, can be from heritable or environmental sources, and are a subset of colony odor. The cuticular hydrocarbons can be used as a model for heritable nestmate recognition cues. We propose that because potential nestmate recognition cues, both environmental and genetic, are dynamic in nature rather than static, during its lifetime a worker must continually update its perception (template) of colony odor and nestmate recognition cues.     

Identification of ant repellent allomone produced by social waspPolistes fuscatus (Hymenoptera: Vespidae)

Two active components were isolated from the ant repellent secretion produced by glandular cells on the sixth (terminal) gastral sternite ofPolistes fuscatus (F.) females. One of the components was identified as methyl palmitate. Field bioassays with synthetic methyl palmitate showed it had repellent activity againstSolenopsis geminata (F.)Forelius pruinosus (Roger), andPheidole sp. The effectiveness of the repellent was greater againstF. pruinosus andPheidole sp. than againstS. geminata. Four methyl ester homologs of methyl palmitate also were field tested for repellent activity: methyl myristate had repellent activity against all three ant species, whereas methyl stearate, methyl linoleate, and methyl linolenate exhibited no repellency.     

Interspecific recognition among termites of the genusReticulitermes: Evidence for a role for the cuticular hydrocarbons

Two species of termites,Reticulitermes (lucifugus) grassei andR. (l.) banyulensis, show a high degree of aggressivity toward each other. The epicuticular signature, recognized by contact, can be extracted using organic solvents, and the removal of the signature abolished all types of aggressive behavior. The signature can be transferred to lures, where it triggers interspecies aggression. It was found to be mainly present in the apolar fraction of the cuticular extracts, which contained only hydrocarbons, are determined by GC/MS techniques. Chemical recognition contributes towards isolation of the two species belonging to theR. lucifugus complex.     

Chemical analysis of sternal gland secretion of paper waspPolistes dominulus (Christ) and its social parasitePolistes sulcifer (Zimmermann) (Hymenoptera: Vespidae)

The secretion of the tegumental glands of the fifth and sixth gastral sternites ofP. dominulus and of its social parasiteP. sulcifer was analyzed by gas chromatography-mass spectrometry. Seven long-chain carboxylic acids have been identified: hexadecanoic, octadecadienoic and octadecenoic acids are the major components of the gland secretion ofP. dominulus and octadecenoic acid the main compounds ofP. sulcifer sternal gland secretion. The same carboxylic acids have been found on the nest pedicel.     

Social closure,aggressive behavior,and cuticular hydrocarbon profiles in the polydomous antCataglyphis iberica (hymenoptera,Formicidae)

Nestmate recognition was studied in the polydomous antCataglyphis iberica (Formicinae) in the laboratory. The study examined six colonies collected from two different populations 600 km apart in the Iberian peninsula (Barcelona and Murcia). Introduction of an alien worker into an allocolonial arena always ended in death to the intruder, demonstrating that in this species societies are extremely closed. Dyadic encounters composed of individuals from different colonies in a neutral arena confirmed the existence of high aggression between allocolonial individuals. We also investigated variability in the composition of the major cuticular hydrocarbons between the colonies used in the behaviorial tests. There were marked quantitative differences between the profiles of ants from the two populations, suggesting that the populations are completely segregated. Cuticular profiles within a population tended to be more similar, but were nevertheless colony specific. The degree of colony closure inC. iberica seemed to be independent of geographic distance since aggression between the colonies was always at its maximum, irrespective of their population origin.     

Mechanism underlying cuticular hydrocarbon homogeneity in the antCamponotus vagus (SCOP.) (Hymenoptera: Formicidae): Role of postpharyngeal glands

The aim of the present study was to investigate the regulation and the homogeneity of the chemical signature between members ofCamponotus vagus after experimentally changing the cuticular chemical signature by topically applying hydrocarbons. Topical application of pentane (solvent) to the cuticle of isolated workers led to a significant decrease in the quantities of the cuticular hydrocarbons measured within 3 hr, followed by an increase within the following 3 hr and a period of relative stability from 9 hr to 14 days. On the other hand, after topical application to isolated workers ofn-tetracosane, a hydrocarbon existing only in trace quantity in this species, the quantity of this hydrocarbon measured over time in the epicuticular wax tended to level out at about 14 days after treatment. In contrast, topically applied (Z)-9-tricosene, an unsaturated hydrocarbon not normally synthesized by this species, decreased dramatically within a few hours and had completely disappeared within 14 days. (Z)-9-Tricosene applied to one member of a group was present in the postpharyngeal glands of the other members from 30 min to seven days of cohabitation. The highest levels were recorded in all six workers in each group after one day. GC-MS analyses showed that (Z)-9-tricosene was present in the cuticles of some untreated workers only after four and seven days of cohabitation with a treated worker. These data suggest: (1) that the deposited (Z)-9-tricosene decreased very quickly on the cuticle of the treated worker, although the total amount was spread over the cuticle and postpharyngeal gland and (2) that it was absorbed by the nontreated workers via the postpharyngeal glands during licking or grooming activities and reincorporated into the cuticle at four and seven days. When the treated worker was separated from the other ants by a wire mesh, (Z)-9-tricosene was detected neither in the cuticle nor in the postpharyngeal gland of nontreated workers.     

Mediation of host selection by cuticular hydrocarbons in the honeybee tracheal MiteAcarapis woodi (Rennie)

Using a simple two-choice bioassay and video analysis of individual locomotory tracks, it was determined that a preference for young-bee hosts over old-bee hosts in female honeybee tracheal mites,Acarapis woodi (Rennie), is chemically mediated. When presented with a choice of cuticular extracts from 5-day-old and <1-day-old adult bees, mites showed a significant preference for the young-bee extract in three of four bee colony sources. This discrimination was due apparently to a greater positive response elicited by the young-bee extract rather than a deterrent effect of old-bee extract, as the extract of old bees evoked a significantly higher response than a hexane control. A bioassay of silica-gel fractions of young-bee extract indicated that cuticular hydrocarbons alone were responsible for tracheal mite response. Further fractionation of a hydrocarbon fraction by argentation chromatography demonstrated that both saturated and unsaturated hydrocarbons were involved in the mite response, but when presented in opposition, mites showed a stronger response to the saturated than to the unsaturated components. Mites placed in zones treated with cuticular extract of young bees exhibited higher angular velocities than those placed on hexane, causing them to remain in the extract-treated zones for extended periods. These results point to a possible control strategy whereby migration of tracheal mites to young-bee hosts could be disrupted by artificially altering the chemical profile of the honeybee cuticle.     

Role of cuticular hydrocarbons of aphid parasitoids in their relationship to aphid-attending ants

Lysiphlebus cardui, the dominant aphidiid parasitoid of the black bean aphid,Aphis fabae cirsiiacanthoidis (Afc), on creeping thistle, is able to forage in ant-attended aphid colonies without being attacked by ants. Several behavioral observations and experimental studies led to the hypothesis thatL. cardui mimics the cuticular hydrocarbon profile of its host aphid. Chemical analysis of the cuticular extracts revealed that bothL. cardui and Afc exclusively possess saturated hydrocarbons:n-alkanes, monomethyl (MMA), dimethyl (DMA), and trimethyl alkanes (TMA). Comparison of the hydrocarbon profiles of parasitoid and aphid showed great qualitative resemblance between parasitoid and host:L. cardui possesses almost all host-specific compounds in addition to species-specific hydrocarbons of mainly higher molecular weight (>C₃₀). However, there is a lesser quantitative correspondence between parasitoid and host aphid. Furthermore, we analyzed the cuticular hydrocarbon profile of another parasitoid of Afc,Trioxys angelicae. This aphidiid species is vigorously attacked and finally killed by honeydewcollecting ants when encountered in aphid colonies. Its cuticular hydrocarbon profile is characterized by the presence of large amounts of (Z)-11-alkenes of chain lenghts C₂₇, C₂₉, C₃₁, and C₃₃, in addition to alkanes and presumably trienes. The role of the unsaturated hydrocarbons onT. angelicae as recognition cues for aphid-attending ants is discussed.     

Cuticular hydrocarbons wherebyMessor barbarus ant workers putatively discriminate between monogynous and polygynous colonies. Are workers labeled by queens?

The results of laboratory experiments carried out with both monogynous and artificially polygynousMessor barbarus ant colonies (which under natural conditions are always monogynous) have shown that the workers belonging to monogynous colonies were able to discriminate between intruders from other monogynous colonies and those from polygynous (di- and trigynous) ones. What mechanisms are involved in this discriminatory ability? Since differences in the relative proportions of the hydrocarbons they carry are known to convey complex messages that are used for recognition purposes, it was proposed here to investigate whether there existed any differences in the proportions of the various hydrocarbons carried by the diverse categories of intruders tested in our experiments. It emerged that one set of hydrocarbons, which were usually present in rather small proportions and included all the families that constitute this species' chemical signature (n-alkanes, mono-, di-, and trimethylalkanes), was characteristically associated with workers from monogynous colonies. Another set of hydrocarbons, which included some of the above components, mostly in larger relative proportions, can be said to have characterized the digynous and trigynous colonies.     

Effects of relative humidity,temperature, and population density on production of cuticular hydrocarbons in housefly Musca domestica L

The production of cuticular hydrocarbons by both males and females of Musca domestica L. under very wet conditions (90% relative humidity) compared to the production at 50 and 20% relative humidity is delayed up to at least 3 days after emergence from the pupae. Eight days after emergence, however, males contain the same amounts of hydrocarbons at 90, 50, and 20% relative humidity, whereas females at 90% still possess less of these substances than at 50 and 20%. It is suggested that this is due to the fact that males, being more active than females, need more cuticular hydrocarbons to prevent water loss. No indication is found that relative humidity has a different effect on the production of the sex pheromone, muscalure [(Z)-9-tricosene] by females than on the production of the other hydrocarbons. Male and female flies produce more hydrocarbons at 35°C than at 20°C. On females, the relative amounts of nonacosane and methyl- and dimethylnonacosanes are significantly higher at 35°C than at 20°C. Female flies produce some (Z)-9-tricosene after eight generations at low population density in contrast to females at high population density, which did not produce muscalure. We suggest that because of the relatively large contribution to the total population, the properties of a small number of females are likely to be expressed sooner in the next generations of small populations than in those of large populations.     

Novel internally branched,internal alkenes as major components of the cuticular hydrocarbons of the primitive australian antNothomyrmecia macrops Clark (Hymenoptera: Formicidae)

Internally mono- and dimethyl branched, internal alkenes, which constitute most of the cuticular hydrocarbon present on workers of the primitive Australian antNothomyrmecia macrops Clark, have been identified by gas chromatography-mass spectrometry. They are the first such alkenes reported from insects. Also present are alkanes with similar carbon skeletons. The hydrocarbon patterns of three separateNothomyrmecia colonies were very similar.     

Hydrocarbons ofNasutitermes acajutlae and comparison of methodologies for sampling cuticular hydrocarbons of caribbean termites for taxonomic and ecological studies

Using data from the arboreal nestingNasutitermes acajutlae (Holmgren), we propose standard collection and extraction methodology for characterization of cuticular hydrocarbons of termites under field conditions in the tropics. Specifically, we evaluated: (1) the effect of the duration and the number of extractions; (2) the effect of drying termites before extraction; (3) the effect of sample size; (4) the effect of solvents (ethanol versus hexane) on cuticular hydrocarbon profiles. Olefins comprise ca. 70% of the cuticular hydrocarbons ofN. acajutlae. Hydrocarbons consist of two distinct groups: early-eluting components, primarilyn-alkanes and methyl-branched alkanes, and late-eluting compounds, which consist almost exclusively of unsaturated components with one to six double bonds. Soldiers have more early-eluting compounds than workers or alates. Nests from the same island had qualitatively similar, but quantitatively dissimilar hydrocarbon mixtures. Brief extractions of 300 live workers in 10 ml of hexane for only 20 sec produced a hydrocarbon mixture equivalent to a 10-min extraction. Long-term extraction of 300 workers in hexane for two years resulted in different mixtures of hydrocarbons. Drying workers tended to enhance extraction of the less abundant unsaturated compounds such as C_41.4 and C_41.5. A single extraction of a minimum of 100 workers (live or dried), with hexane for 20 sec to 10 min is best; these extraction regimes resulted in mixtures of hydrocarbons that are quantitatively very similar. For quantitative comparisons, extracts from dried samples should not be compared to those from live samples. Storage in ethanol caused numerous unidentified, nonhydrocarbon compounds to be extracted either from the cuticle or from internal tissues.