Hydrocarbon dynamics within and between nestmates inCataglyphis niger (Hymenoptera: Formicidae)

The objective of the present study was to evaluate the interrelationship between the cuticular and postpharyngeal glands' hydrocarbons, both in the individual ant and during its interaction with nestmates. In vivo radiochemical assays were employed to monitor the de novo hydrocarbon biosynthesis from acetate in the ant's body. The newly synthesized hydrocarbons appeared first internally and after 24 hr they accumulated in the postpharyngeal gland and on the cuticular surface. Blocking the possibility of external transfer of hydrocarbons between cuticle and postpharyngeal gland led to a significant decrease of labeled hydrocarbons in the postpharyngeal gland. In addition, during encounters between labeled and unlabeled ants, newly synthesized hydrocarbons were transferred, mainly via trophallaxis, but also by allo-grooming and physical contact. In view of these results, we propose as a model for their dynamics that hydrocarbons are synthesized in tissues associated with the integument. Through self-grooming, there is a constant exchange of hydrocarbons between the cuticular surface and the postpharyngeal gland. Furthermore, in encounters between nestmates, hydrocarbons are exchanged among them mostly by trophallaxis, with the mediation of the postpharyngeal gland. Thus, this gland acts as a pool for mixing colonial hydrocarbons and may serve to attain a unified colony odor.     

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.     

Termite predation byMegaponera foetens (FAB.) (Hymenoptera: Formicidae)

Termite predation by the ponerine ant,Megaponera foetens, is coordinated by chemicals from at least two glands. Columns of ants are guided to termite foraging areas by pheromones originating from the poison apparatus. On finding groups of termites, ants release alkyl sulfides (dimethyl disulfide and trisulfide) from their mandibular glands which attract sister workers who dig, into the termite galleries, in response to other unidentified mandibular gland pheromones.     

Explanation of Bitter Taste of Venom of Ponerine Ant, Pachycondyla apicalis

The venom gland of workers of Pachycondyla (= Neoponera) apicalis (Hymenoptera: Formicidae) contains the bitter-tasting cyclic dipeptide of leucine and phenylalanine [cyclo-leu-phe or 3-benzyl-6-(2-methylpropyl)-2,5-piperazinedione]. The venom also contains proteins of undetermined activity. It is suggested that the function of the venom may be both defensive and offensive. The mandibular glands of N. apicalis contains δ-decalactone and benzaldehyde.     

Food Wrapping with the Postpharyngeal Gland Secretion by Females of the European beewolf <Emphasis Type="Italic">Philanthus triangulum</Emphasis>

Ground-nesting animals share their habitat with countless microorganisms that can play important roles as pathogens or competitors for food resources. Thus, species that store food in the soil, either for themselves or for their progeny, must protect these resources against microbial degradation. Females of the European beewolf, Philanthus triangulum, hunt honeybees as provisions for their brood and store the paralyzed prey in their subterranean nests. A previous study had shown that females lick the surface of prey before oviposition and that this licking treatment delays mold growth. Here, we showed that females apply large amounts of a secretion from their postpharyngeal glands onto the surface of their prey during the licking behavior. Inhibition-zone assays showed that comparatively large amounts of the gland secretion had no direct antimycotic effect. We discuss our findings with regard to other possible mechanisms of the postpharyngeal gland secretion against fungal growth.     

How an Ant Manages to Display Individual and Colonial Signals by Using the Same Channel

Cuticular hydrocarbons are used by some ants to discriminate nestmates from nonnestmates. Every member of the colony bears the same pattern because they are continuously exchanged among nestmates. The postpharyngeal gland (PPG) stores the blend of hydrocarbons and is involved in the distribution of this common mixture. However, some individuals might display individual information on the cuticle (such as a chemical signal of fertility) that must not be mixed within the common pool. We investigated how this paradox is solved in the ant Pachycondyla goeldii by analyzing the nature and localization of colonial and fertility signals. Workers in a queenless condition showed a dominance hierarchy that was correlated with ovarian development. Hydrocarbons from the cuticle and the PPG analyzed by gas chromatography (GC) and identified by GC-mass spectrometry showed a clear discrimination among colonies, supporting the involvement of the PPG in the colonial identity signal. We identified and selected 11 cuticular hydrocarbons that permitted us to discriminate ovarian development classes and that might function as a fertility signal. They allowed clear colony discrimination as well, which suggests that the two signals (the individual signal of fertility and the common signal of colony identity) can be conveyed by the same compounds. However, the hydrocarbons in the PPG did not discriminate among ovarian developmental classes, suggesting that the portion of variation in the cuticular hydrocarbons constituting the fertility signal is superimposed on the signal of colony identity.     

Properties of cuticular oxidases used for sex pheromone biosynthesis byHeliothis zea

Biosynthesis of the aldehydic sex pheromone components released by females ofHeliothis zea was found to be catalyzed by primary alcohol oxidases residing in the cuticle that covers the glands. Activity, as indicated by conversion of primary alcohol to aldehyde, was as high in cell-free cuticle as it was in intact pheromone glands. Studies indicated that some activity was associated with the surface of the epicuticle and could be removed, into buffer, by sonication. However, the majority of activity lies within the inner epicuticle and exo- and endocuticular layers. The oxidase was not functional in pharate pupae that did not have mature adult cuticle but became functional just prior to adult emergence. The enzyme in individual glands was saturated at alcohol concentrations above 100 n. moles. Nonionic detergents did not affect the activity of the oxidase in the cuticle but treatment with either 7 M urea or 1% SDS resulted in total loss of activity. Studies on the effect of pH indicated an optimum at 6.4; however, activity was high throughout the range of 5–9. The oxidase was functional in both dichloromethane and hexane, suggesting that this enzyme system may have applications for organic synthesis of aldehydes.     

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.     

Species Recognition from Postpharyngeal Gland Contents of Ants of the Cataglyphis bicolor Group

The Cataglyphis bicolor group of species of desert-dwelling ants, difficult to identify from morphological features alone, can be readily recognized by the contents of their postpharyngeal glands. Analysis by linked gas chromatography–mass spectrometry of glands from colonies identified only by code numbers showed in all samples straight and branched-chain alkanes and linear alkenes. C. viaticus, C. bicolor, and C. savignyi, the three species most difficult to distinguish morphologically, each contained distinctly different patterns of hydrocarbons, as illustrated by cluster analysis. The 16 most abundant hydrocarbons in the whole group of samples were selected and plotted as windrose diagrams. The differences in the windroses have more visual impact than gas chromatograms of the same data. The only case where there was any similarity was that between C. bicolor and C. diehlii, and even there the resemblance was not close. C. bombycinus is a sympatric species but is recognized as not belonging to the bicolor group by its different mandibular gland substances. It also was easily distinguished by its postpharyngeal gland contents from the other species.     

A Non-lethal Water-based Removal-reapplication Technique for Behavioral Analysis of Cuticular Compounds of Ants

Interspecific relationships among insects are often mediated by chemical cues, including non-volatile cuticular compounds. Most of these compounds are hydrocarbons that necessitate the use of solvents for their extraction, identification, and manipulation during behavioral assays. The toxicity of these solvents often precludes the removal and reapplication of hydrocarbons from and to live insects. As a consequence, dummies often are used in behavioral assays, but their passivity can bias the behavior of the responding insects. To overcome these limitations, we propose a method where cuticular compounds are extracted from live ants by placing them into glass vials half-filled with tepid water (ca. 34°C) and vigorously shaking the vials to form an emulsion whose supernatant can be analyzed and/or reapplied to other ants. We demonstrate that cuticular compounds can be extracted from workers of the red fire ant, Solenopsis saevissima, and reapplied to the cuticle of workers from a sympatric species, Camponotus blandus (both Hymenoptera: Formicidae), while keeping the ants alive. Gas chromatographic-mass spectrometric analysis and behavioral assays were used to confirm the successful transfer of the behaviorally active compounds.     

Chemical Ecology of the Defense of Two Nymphalid Butterfly Larvae Against Ants

We analyzed the behavioral responses of the ants Camponotus rufipes and Solenopsis geminata towards all instars of Dione junio and Abananote hylonome. We also analyzed ant behavior towards hexane extracts of larvae and extracts of the spines and neck glands of the fifth instars of both species and identified the chemical compounds present. Larvae of both species were repellent to ants from the first instar onward. Later instars survived ant attacks better than earlier instars. The spines and neck glands of the larvae influenced the behavior of C. rufipes. The chemical compounds contained in the hexane extracts of whole first and fifth instars and in the spines and neck glands of fifth instars were principally carboxylic acids and terpenes. Further bioassays confirmed the repellent effect of some of these acids toward ants.     

Neotropical ant gardens

In ant gardens of lowland Amazonia, parabiotic ant speciesCamponotus femoratus andCrematogaster cf.limata parabiotica cultivate a taxonomically diverse group of epiphytic plants, whose establishment is restricted to arboreal carton ant nests. Epiphyte seeds are collected by workers ofCa. femoratus, the larger of the two ants, and stored unharmed in brood chambers where they subsequently germinate. Although seeds of some ant-garden epiphytes bear nutritional rewards, previous studies have shown that these rewards are not sufficient to explain the pattern of ant attraction to seeds. Five aromatic compounds occur frequently in and on the seeds of most ant-garden epiphytes and may be chemical cues by which ants recognize propagules of their symbiotic plants. The most widely distributed of these is methyl 6-methylsalicylate [6-MMS]1, previously reported as a major mandibular gland product in relatedCamponotus species and present in trace quantities inCa. femoratus males. (–)-Citronellol6 (previously unreported inCamponotus) was the principal volatile constituent in extracts of male heads, and (–)-mellein7 was present in small quantities. Discovery of 6-MMS inside the mandibular glands of maleCa. femoratus (and its presence in analogous glands of related ants) offers preliminary support for Ule's (1906) hypothesis that seeds attract ants by mimicking ant brood. In addition, the likely fungistatic activity of seed compounds suggests that they could retard microbial pathogens of ants and plants in the organic detritus of nest gardens. While the presence of identical seed compounds in so many unrelated plant lineages might represent a remarkable case of convergent evolution, other interpretations are possible.     

Similarity of Cuticular Lipids Between a Caterpillar and Its Host Plant: A Way to Make Prey Undetectable for Predatory Ants?

Ithomiine butterflies (Nymphalidae) have long-lived, aposematic, chemically protected adults. However, little is known about the defense mechanisms in larvae and other juvenile stages. We showed that larvae Mechanitis polymnia are defended from ants by a chemical similarity between their cuticular lipids and those of the host plant, Solanum tabacifolium (Solanaceae). This is a novel defense mechanism in phytophagous insects. A field survey during one season showed that larval survivorship was up to 80%, which is high when compared with other juvenile stages. In a laboratory bioassay, live larvae on their host plant were not attacked by the predatory ant Camponotus crassus (Formicidae). Two experiments showed that the similarity between the cuticular lipids of M. polymnia and S. tabacifolium protected the larvae from C. crassus: (a) when the caterpillar was switched from a host plant to a non-host plant, the predation rate increased, and (b) when a palatable larva (Spodoptera frugiperda, Noctuidae) was coated with the cuticular lipids of M. polymnia and placed on S. tabacifolium leaves, it no longer experienced a high predation rate. This defensive mechanism can be defined as chemical camouflage, and may have a double adaptive advantage, namely, protection against predation and a reduction in the cost of sequestering toxic compounds from the host plant.     

Task-Related Environment Alters the Cuticular Hydrocarbon Composition of Harvester Ants

Within a colony of harvester ants (Pogonomyrmex barbatus), workers in different task groups differ in the hydrocarbon composition of the cuticle. Foragers and patrollers, which spend extended periods of time outside the nest, have a higher proportion of saturated, unbranched hydrocarbons (n-alkanes) on the cuticle than nest maintenance workers, which spend only short periods of time outside the nest. We tested whether these task-related differences in ant cuticular chemistry arise from exposure to conditions outside the nest. Nest maintenance workers experiencing daily, short-term outside exposure developed a higher proportion of n-alkanes on the cuticle than workers kept inside the lab. Independent manipulations of ultraviolet radiation, relative humidity, and temperature revealed that only the combination of high temperature (ca. 38°C) and low relative humidity (ca. 8%) increased the proportion of cuticular n-alkanes. The results indicate that warm dry conditions, such as those encountered when an ant leaves the nest, trigger changes in cuticular chemistry.     

Ant repellent effect of the sternal gland secretion ofPolistes dominulus (Christ) andP. sulcifer (Zimmermann). (Hymenoptera: Vespidae)

The long-chain carboxylic acids identified in the sternal gland secretion ofPolistes dominulus andP. sulcifer females were tested individually on three species of ants,Crematogaster scutellaris, Formica cunicularia, andLasius sp., in order to verify if they have a repellent effect. The unsaturated acids (palmitoleic, linoleic, and oleic) act as repellents of all three ant species, while the saturated acids (lauric, myristic, palmitic, and stearic) have no effect. The mixture reproducing the secretion of the sternal glands ofP. dominulus maintained its repellency for at least four days.     

Comparative survey of abdominal gland secretions of the ant subfamily Ponerinae

The chemical contents of three abdominal glands were investigated in representative species of the ponerine ants. The Dufour glands of 14 species show a wide variety of contents. In Mystrium camillae and Proceratium itoi, no volatile substances were found in either the Dufour or venom glands. In Ectatomma sp., Diacamma ceylonense, Diacamma indicum, Pachycondyla obscuricornis, and Pachycondyla striata, volatile chemicals were found in the venom glands as well as in the Dufour glands. Platythyrea punctata was examined, but unusually it does not have a Dufour gland and its venom gland contained no volatile substances. Epoxides were found in ants for the first time in the Dufour glands of Amblyopone reclinata. Venom glands of Pachycondyla tarsata were also found to contain volatile material, including bitter-tasting cyclic dipeptides. In all, 16 species have been added to the list of those examined. All of the 27 known analyses of Dufour glands, 21 analyses of venom glands, and 4 of pygidial glands of workers of ponerine ant species have been brought together in order to seek some pattern in the type of glandular contents. Although the great majority of species produce hydrocarbons in their worker Dufour glands, and some have terpenes, there is no observable pattern for this gland on a tribe or genus level. Volatile compounds have been found in the venom glands of some species of the tribe Ponerini only. The information on pygidial glands is still too fragmentary for any conclusions.     

Antennal Olfactory Physiology and Behavior of Males of the Ponerine Ant <Emphasis Type="Italic">Harpegnathos saltator</Emphasis>

In comparison to the large amount of study on the communication abilities of females in ant societies and their associated chemical ecology and sensory physiology, such study of male ants has been largely ignored; accordingly, little is known about their olfactory sensory capabilities. To address this, we explored peripheral odor sensitivities in male Harpegnathos saltator by measuring the electrophysiological activity of olfactory sensory neurons within antennal trichoid and coeloconic sensilla using an extracellular recording technique. In an initial trial of 46 compounds, sensilla trichodea responded strongly to two alarm pheromone components, while a limited number of non-hydrocarbon odorants elicited strong responses in sensilla coeloconica. Both sensillar types responded indifferently to 31 cuticular hydrocarbons (CHCs) and synthetic long-chain hydrocarbons (HCs) typically found on insect cuticle. In a search for sensilla responding to CHCs and other compounds, we found some sensilla that responded to synthetic HCs and CHCs from virgin queen postpharyngeal glands that are potentially used in close range mate recognition. Olfactometer bioassays of male ants to 15 non-HCs correlated sensory responsiveness to the respective behavioral responses. Comparing olfactory responses between H. saltator males and females, we found that sensilla coeloconica and basiconica of workers showed greater responses and broader selectivity to all compounds. The rarity of CHC-responding trichoid sensilla in Harpegnathos males suggests a more specific role in sexual communication compared to that in females, which use CHCs in a broader communication context.     

Biochemical and behavioral evidence foe hybridization between fire ants,Solenopsis invicta andSolenopsis richteri (Hymenoptera: Formicidae)

Behavioral and biochemical evidence is presented for hybridization between the fire ants,Solenopsis richteri andS. invicta. The response of the two species to extracts of their trail pheromones presented as a point source is clearly species-specific; however, hybrid workers responded to parental Dufour's gland extracts and parental workers responded to Dufour's gland extracts of the hybrid. The behavioral evidence for hybridization was confirmed by gas Chromatograph comparison of the Dufour's gland extracts of the three fire ant forms, which showed a pattern for the hybrid that was intermediate to the two parental species.     

Solitary Foraging in the Ancestral South American Ant, <Emphasis Type="Italic">Pogonomyrmex vermiculatus</Emphasis>. Is it Due to Constraints in the Production or Perception of Trail Pheromones?

Several North American species of Pogonomyrmex harvester ants exhibit group foraging, whereas South American species are exclusively solitary foragers. The composition of the secretions of the poison and Dufour glands in the South American species, Pogonomyrmex vermiculatus, were analyzed, and the secretions and their components were tested as trail pheromones in laboratory bioassays. The major compounds in the poison gland were the alkylpyrazines, 2,5-dimethylpyrazine, 2,3,5-trimethylpyrazine, and 3-ethyl-2,5-dimethylpyrazine. The Dufour gland contained five alkanes, from tridecane to heptadecane, with pentadecane being most abundant. In behavioral bioassays, poison gland extracts and the mixture of pyrazines produced a trail pheromone effect, whereas the Dufour gland extracts and the alkanes had no effect on ant locomotion. We conclude that group foraging in P. vermiculatus does not arise from the inability to produce or detect possible pheromones, but rather, from physiological and/or ecological factors.     

Fine Tuning of Social Integration by Two Myrmecophiles of the Ponerine Army Ant, <Emphasis Type="Italic">Leptogenys distinguenda</Emphasis>

Myrmecophiles are animals that live in close association with ants and that frequently develop elaborate mechanisms to infiltrate their well-defended host societies. We compare the social integration strategies of two myrmecophilic species, the spider, Gamasomorpha maschwitzi, and the newly described silverfish, Malayatelura ponerophila gen. n. sp. n., into colonies of the ponerine army ant, Leptogenys distinguenda (Emery) (Hymenoptera: Formicidae). Both symbionts use chemical mimicry through adoption of host cuticular hydrocarbons. Exchange experiments between L. distinguenda and an undetermined Leptogenys species demonstrate that reduced aggression toward alien ants and increased social acceptance occurred with individuals of higher chemical similarity in their cuticular hydrocarbon profiles. We found striking differences in chemical and behavioral strategies between the two myrmecophiles. Spider cuticular hydrocarbon profiles were chemically less similar to the host than silverfish profiles were. Nevertheless, spiders received significantly fewer attacks from host ants and survived longer in laboratory colonies, whereas silverfish were treated with high aggression and were killed more frequently. When discovered and confronted by the host, silverfish tended to escape and were chased aggressively, whereas spiders remained in contact with the confronting host ant until aggression ceased. Thus, spiders relied less on chemical mimicry but were nevertheless accepted more frequently by the host on the basis of behavioral mechanisms. These findings give insights into the fine tuning of social integration mechanisms and show the significance of qualitative differences among strategies. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.