Hydrocarbon site of synthesis and circulation in the desert ant Cataglyphis niger

Chemical analyses revealed that in Cataglyphis niger both the hemolymph and the crop contain the same hydrocarbons that are found in the postpharyngeal gland (PPG). On the cuticle, on the other hand, alkanes, and in particular nonacosane, were more abundant than in the PPG. Studies of their biosynthesis in vivo, using intact ants, revealed the presence of newly synthesized hydrocarbons in both the PPG and the crop. In decapitated ants (in the absence of the PPG), however, the crop did not contain any newly synthesized hydrocarbons, indicating the PPG as the major source of crop hydrocarbons. The fat body, as demonstrated by in vitro studies, is the major tissue that biosynthesizes hydrocarbons. The PPG failed to do so, but showed good de novo biosynthesis of other lipid constituents. The large amount of hydrocarbons in the crop suggests that the alimentary canal may serve as an outlet for the overflow of PPG hydrocarbons, or as a route for the directed clearance of hydrocarbons from the PPG.These results confirm and enlarge the model proposed for hydrocarbon circulation in C. niger. They are synthesized by the fat body, released to the hemolymph and transported to the cuticle and the PPG. The PPG hydrocarbons are applied to the cuticle by self-grooming, but can also be cleared via the alimentary canal. Partial emptying of the PPG may facilitate the admixing of recognition cues that the ant may acquire from nestmates by trophallaxis. The reason for the dissimilarity in hydrocarbon composition between the PPG and the cuticle is not yet clear; it may be due to secretions from additional glands, or reflect deviant hydrocarbon transport mechanisms between the PPG and the cuticle.     

Is the postpharyngeal gland of a solitary digger wasp homologous to ants? Evidence from chemistry and physiology

The postpharyngeal gland (PPG) was thought to be restricted to ants where it serves a crucial function in the generation of the colony odour. Recently, head glands that closely resemble the PPG of ants were discovered in females of a solitary digger wasp, the European beewolf. The function of this gland necessarily differs from ants: beewolf females apply the secretion of their PPG onto the bodies of paralysed honeybees that serve as larval provisions in order to delay fungus growth. Since ants and digger wasps are not closely related, the occurrence of this gland in these two taxa might either be due to convergent evolution or it is a homologous organ inherited from a common ancestor. Here we test the hypothesis that the PPGs of both taxa are homologous by comparing characteristics of chemical composition and physiology of the PPG of beewolves and ants. Based on reported characteristics of the PPG content of ants, we tested three predictions that were all met. First, the PPG of beewolves contained mainly long-chain hydrocarbons and very few compounds with functional groups. Second, the composition of hydrocarbons in the beewolf PPG was similar to that of the hemolymph. Taking the structure of the gland epithelium and the huge requirements of beewolf females for gland secretion into account this result suggests that the content of the PPG is also sequestered from the hemolymph in beewolves. Third, the chemical composition of the PPG and the cuticle was similar in beewolves since cuticular hydrocarbons derive either from the hemolymph or the PPG. Taking the considerable morphological similarities into account, our results support the hypothesis of a homologous origin of the PPG in beewolves and ants.     

Task-related variation of postpharyngeal and cuticular hydrocarbon compositions in the ant Myrmicaria eumenoides

In the ant Myrmicaria eumenoides we investigated postpharyngeal and cuticular hydrocarbons. At eclosion the glands contained almost no hydrocarbons and there were no lipid inclusions in the glandular epithelium. During the first 3 weeks of adult life the amount of hydrocarbons in the gland increased until day 5, and then remained constant while the lipid content in the epithelium increased steadily. Intracolonial hydrocarbon compositions were not uniform. Compositions of post-pharyngeal and cuticular hydrocarbons in individual ants varied simultaneously, but in different manner depending on the tasks of the ant (brood-tenders, foragers, scouts). Variations on the cuticle were greater than in the gland, but they were strongly correlated. Independent of ants' age and task, cuticular hydrocarbon compositions were dominated by alkenes and alkadienes. Task-specific differences in cuticular compositions were mainly in the amount of alkenes (high in foragers) and alkadienes (high in brood-tenders). Variation of hydrocarbons was low in ants up to 10 weeks old. Thereafter, ants fell into two groups: (1) ants that did not change their hydrocarbons and remained in the nest, and (2) ants that changed their hydrocarbon compositions and became foragers. These results contribute to an ongoing discussion of the dynamic relationship between post-pharyngeal and cuticular hydrocarbons.     

Structure, chemical composition and putative function of the postpharyngeal gland of the emerald cockroach wasp, Ampulex compressa (Hymenoptera, Ampulicidae)

The postpharyngeal gland (PPG) plays a major role in the social integration of ant colonies. It had been thought to be restricted to ants but was recently also described for a solitary wasp, the European beewolf (Philanthus triangulum). This finding posed the question whether the gland has evolved independently in the two taxa or has been inherited from a common ancestor and is hence homologous. The latter alternative would be supported if a PPG was found in more basal taxa. Therefore, we examined a species at the base of the Apoidea, the solitary ampulicid wasp Ampulex compressa, for the existence of a PPG. Both sexes of this species possess a cephalic gland that branches off the posterior part of the pharynx, is lined by a cuticular intima and surrounded by a monolayered epithelium with the epithelial cells bearing long hairs. Most of these morphological characteristics conform to those of the PPG of ants and beewolves. Chemical analysis of the gland content revealed that it contains mainly hydrocarbons and that there is a congruence of the pattern of hydrocarbons in the gland, on the cuticle, and in the hemolymph, as has also been reported for both ants and beewolves. Based on these morphological and chemical results we propose that the newly described cephalic gland is a PPG and discuss its possible function in A. compressa. The present study supports the view of a homologous origin of the PPG in the aculeate Hymenoptera.     

Ontogeny of hydrocarbon profiles in the ant Aphaenogaster senilis and effects of social isolation

In ants, cuticular hydrocarbons are used for nestmate recognition; they are stored in the postpharyngeal gland and shared among the individuals. Newly emerged ants have a very small quantity of hydrocarbons. We studied the ontogeny of the hydrocarbon profile in Aphaenogaster senilis. The total quantities of both cuticular and postpharyngeal gland (PPG) hydrocarbons increased with age from 0 to 20 days after emergence and then stabilised. These quantities are correlated with the development of the ovary. Under individual social isolation, cuticular hydrocarbons increased as normal, but the total quantity of PPG hydrocarbons never increased from the initial low level. This effect of social isolation on the PPG hydrocarbon level indicates the importance of hydrocarbon transfer between nestmates through the PPG and lends support to the gestalt model of nestmate recognition. To cite this article: K. Ichinose, A. Lenoir, C. R. Biologies 332 (2009).     

Hydrocarbon circulation and colonial signature in Pachycondyla villosa

In ants, both cuticular and postpharyngeal gland (PPG) hydrocarbons (HCs) have been involved in nestmate recognition. However, no detailed comparison is available. A comparative study including also high density lipophorin (HDLp), an internal HC carrier, was therefore undertaken on Pachycondyla villosa. Purified HDLp is an 820 kDa lipoprotein with a density of 1.114 g/ml and two 245 and 80 kDa apo-proteins. Its hydrocarbon profile is very similar with the cuticular one, in agreement with its hydrocarbon carrier function. Conversely, n-alkanes and externally branched monomethylalkanes are markedly decreased in the PPG. According to their physical properties, this suggests that they are involved in waterproofing on the cuticle. The PPG actually contains only internally branched mono-, dimethylalkanes or monomethylalkenes; their greater fluidity is more adequate for chemical communication. The percentages of some of them are statistically not different between the cuticle and PPG. Their mixtures vary with colonies and they may thus be involved in colonial signature. A scheme for hydrocarbon circulation is discussed, involving lipophorin, cuticle, PPG and self-grooming in one individual, a pathway complementary or alternative to the selective delivery by lipophorin in some other insects. HCs are then distributed between nestmates' cuticles through allo-grooming and physical contacts.     

Postpharyngeal gland secretion as a modifier of aggressive behavior in the myrmicine antManica rubida

The role of postpharyngeal gland secretion in nestmate recognition was tested inManica rubida. Behavioral tests comprised two consecutive encounters between five ants and their color-marked nestmate. The first encounter utilized an untreated ant constituting a control, whereas in the second encounter the marked ant was treated with postpharyngeal gland exudate. Scoring was done using an aggression index obtained by direct observation and an agitation index deduced from a frame-by-frame analysis of videotapes of the various tests. When the glandular secretion originated from an alien ant, the ants became very agitated and were aggressive toward their nestmate. When the exudate originated from a nestmate, the ants generally remained calm, although their rates of self-grooming increased.     

A ‘social’ gland in a solitary wasp? The postpharyngeal gland of female European beewolves (Hymenoptera,Crabronidae)

Exocrine glands play an important role in maintaining the integrity of colonies of social Hymenoptera. The postpharyngeal gland (PPG) of ants is crucial for the generation of a nest odour that enables nestmate recognition. The evolutionary history of this gland is unknown and it was thought to be restricted to ants. Here we describe an exocrine head gland in females of a solitary crabronid wasp, the European beewolf, Philanthus triangulum, that resembles the PPG of ants in many respects. The newly described gland has the same location and the same glove like shape as in ants, and it also has a monolayered epithelium with similar ultrastructure. Unlike in ants, the epithelium bears hairs that reach into the lumen of the gland. Although the PPG of beewolves serves a completely different function it is also associated to an allogrooming behaviour as in ants. Based on these morphological and behavioural similarities as well as similarities in the chemical composition of the content of the PPG of both taxa, we hypothesise that the PPGs of ants and beewolves have a common evolutionary origin. Thus, our results suggest that the PPG in ants might not have evolved in response to social requirements but might have already existed in solitary predecessors.     

Metapleural- and postpharyngeal-gland secretions from workers of the ants Solenopsis invicta and S. geminata

Chemical analyses by GC-MS of the metapleural glands (MG) from workers of Solenopsis invicta and S. geminata revealed for the first time the chemical composition of these glands and showed small differences between the two species. The MG of both species contain oleic, stearic, linoleic, and palmitic acid. Both ants, in addition, have small but significant amounts of hydrocarbons in their MG reservoir, which are the same as those found in their postpharyngeal glands (PPG). The PPG of both species contain alkanes, alkenes, and Me-branched alkanes. Each species is characterized by a specific composition of PPG chemicals with some overlap between species. These results suggest that the MG synthesizes mainly palmitic, linoleic, oleic, and stearic acids in these two ants, whereas PPG contains hydrocarbon mixes that widely vary between these two phylogenetically related species.     

Functional morphology of the postpharyngeal gland of queens and workers of the ant Monomorium pharaonis (L.)

Eelen D., Børgesen L.W. and Billen J. 2006. Functional morphology of the postpharyngeal gland of queens and workers of the ant Monomorium pharaonis (L.). —Acta Zoologica (Stockholm) 87 : 101–111 The postpharyngeal gland (PPG) is unique to ants and is the largest exocrine gland in their head. In queens of the pharaoh's ant, Monomorium pharaonis, the gland contains approximately 15 finger‐like epithelial extensions on each side and opens dorsolaterally in the posterior pharynx. In these ants the PPG morphology varies considerably according to age and mating status. The epithelial thickness increases with age and reaches a maximum at 3 weeks in both virgin and mated queens. A considerable expansion of the lumen diameter occurs in both groups between 4 and 7 days. Virgin queens release their secretion into the gland lumen from an age of 7 days, whereas mated queens accumulate large amounts of secretion in their epithelium. The increasing epithelial thickness, together with the increasing lumen diameter, the presence of numerous inclusions in the epithelium and the release of secretion, are indicative for increasing gland activity. The gland ultrastructure indicates involvement in lipid metabolism and de novo synthesis of lipids. The PPG of workers consists of 12 finger‐like tubes at each side. There is a significant difference in epithelial thickness between nurses and repletes and between nurses and foragers. We suggest the PPG serves different purposes in pharaoh's ants: it is likely that the PPG of workers and virgin queens is used to feed larvae. In mated queens the gland probably plays a role in providing the queen with nutritious oils for egg production. The PPG may also function in signalling species nestmate and caste identity, as well as in the reproductive capacity of the queens.     

Exocrine glands of the ant Myrmoteras iriodum

This paper describes the morphological characteristics of nine major exocrine glands in workers of the formicine ant Myrmoteras iriodum. The elongate mandibles reveal along their entire length a conspicuous intramandibular gland, which contains both class‐1 and class‐3 secretory cells. The secretory cells of the mandibular glands show a peculiar appearance, with a branched end apparatus, which is unusual for ants. The other major glands (pro‐ and postpharyngeal gland, infrabuccal cavity gland, labial gland, metapleural gland, venom gland and Dufour gland) show common features for formicine ants. The precise function of the glands could not yet be experimentally demonstrated, and to clarify this will depend on the availability of live material of these enigmatic ants in future.     

Morphology and ultrastructure of a bacteria cultivation organ: the antennal glands of female European beewolves, Philanthus triangulum (Hymenoptera, Crabronidae)

Females of a solitary digger wasp, the European beewolf (Philanthus triangulum F.), cultivate symbiotic bacteria of the genus Streptomyces in specialized antennal glands. The streptomycetes are secreted in the subterranean brood cells and protect the offspring against mould fungi. We reconstructed the complex morphology of the antennal glands using 3D-visualization software, investigated the ultrastructure of the glands, and examine the role of the antennal glands as organs for the cultivation of the symbiotic bacteria. The bacteria are cultivated in five antennomeres within large reservoirs that consist of two slightly bent lobes. Each gland reservoir is bordered by a monolayered epithelium lined with a partially reinforced cuticle and when completely filled with bacteria it comprises about half of the antennomere's volume. The opening of the reservoir is covered by gelatinous appendage of the cuticle. The cells of the monolayered epithelium bordering each reservoir show basal invaginations, apical microvilli and numerous vesicles. Each reservoir is surrounded by approximately 400 class 3 gland units that are connected to the reservoir lumen through conducting canals. The class 3 gland cells contain numerous vesicles and a high density of rough endoplasmatic reticulum. In the reservoir lumen, large numbers of symbiotic Streptomyces bacteria are embedded in secretion droplets. Thus, the bacteria are apparently provided with large amounts of nutrients via the gland epithelium and the class 3 gland cell units.     

Nest-area marking with faeces: a chemical signature that allows colony-level recognition in seed harvesting ants (Hymenoptera, Formicidae)

Summary. Nestmate recognition systems in ants are largely based on chemical signals. The hydrocarbon fraction of the lipid layer which covers the insect cuticle plays a determinant role in this context. Here we report a novel extension of nestmate and alien recognition – nest area marking with faeces containing the same hydrocarbons as the cuticle of workers – in a harvesting ant, Messor capitatus. Workers of M. capitatus deposit large quantities of brown-yellow material from the hindgut (termed spots) in the vicinity of the nest. Behavioural investigation showed that such spotting behaviour has a communicative value in the context of nest area identification. Anal fluids deposited in the nest surroundings contain colony-specific cues which the ants use to recognize their own nest areas, and distinguish them from foreign areas even in the absence of nestmate or alien ants. Chemical analyses by gas chromatography-mass spectrometry (GC-MS) of the contents of anal spots, rectal sacs, and cuticular extracts revealed that all contain the same long-chained linear and branched hydrocarbons in varying proportions. Importantly, multivariate analyses showed that the relative proportions of these compounds on the cuticle and in spots are colony-specific. This provides a mechanism by which spot marking could be used by workers to define and recognize their colony area, and would represent a simple extension of the existing nestmate recognition template based on colonial cuticular signatures. The ecological and sociobiological implications of these findings are discussed.Received 3 February 2004; revised 10 June 2004; accepted 14 June 2004.     

Morphology,Chemistry and Function of the Postpharyngeal Gland in the South American Digger Wasps Trachypus boharti and Trachypus elongatus

Microbes pose severe threats to animals as competitors or pathogens and strongly affect the evolution of life history traits like parental care. Females of the European beewolf Philanthus triangulum, a solitary digger wasp, provision their offspring with paralyzed honeybees and embalm them with the secretion from large postpharyngeal glands (PPG) that contain mainly unsaturated hydrocarbons. This coating changes the physico-chemical properties of the prey surface, causes a reduction of water condensation and retards growth of mold fungi. Here we examined the closely related South American genus Trachypus, which shows a life-history similar to Philanthus. We investigated whether Trachypus spp. also possess PPGs and embalm larval provisions. Using histological methods and 3D reconstructions we show that Trachypus boharti and T. elongatus possess PPGs that are similar to P. triangulum but somewhat smaller. The ultrastructure of the gland epithelium suggests that the gland content is at least partly sequestered from the hemolymph. Chemical analyses using gas chromatography / mass spectrometry revealed that both the cuticle and PPGs of Trachypus contain mainly unsaturated long-chain hydrocarbons. The gland of T. boharti additionally contains long-chain ketones. The hydrocarbons from the PPG of T. elongatus occurred on prey bees excavated from nests in the field but not on conspecific control bees. While the embalming only slightly elevated the amount of hydrocarbons on prey bees, the proportion of unsaturated hydrocarbons, which is crucial for the antifungal effect, was significantly increased. The Trachypus species under study possess PPGs that are very similar to the PPG of P. triangulum with regard to morphology, ultrastructure and chemistry. Moreover, we provide clear evidence that T. elongatus females embalm their prey, presumably as a means of prey preservation. The observed differences among Trachypus and Philanthus in gland size and prey embalming may have evolved in response to divergent ecological conditions.     

Chemotaxonomy of some Cataglyphis ants from Morocco and Burkina Faso

The taxonomy of Cataglyphis ants has many unsolved problems and chemotaxonomy can provide additional insight for their resolution. We describe here the chemical content of the postpharyngeal and Dufour glands of three Cataglyphis ant species: C. viaticus, C. mauritanicus from Morocco and for the first time a Sub-Saharan Cataglyphis, C. sp. (BF) from Burkina Faso. These three species are very distinct chemically with respect to both the postpharyngeal and Dufour glands. Methyl-alkenes, rare in ants, are characteristic of C. sp. (BF) postpharyngeal glands. A comparison with C. bicolor from Tunisia indicated that C. sp. (BF) can be included into the bicolor group with C. viaticus. We suggest that the content of the Dufour gland is a better phylogenetic indicator than the content of the postpharyngeal gland.     

Dufour gland secretion of Polistes wasp: Chemical composition and possible involvement in nestmate recognition (Hymenoptera: vespidae)

The Dufour gland secretion and cuticle lipids of foundresses of Polistes dominulus have been chemically analysed. The gland secretion (analysed for the first time in a polistine species) contains a complex mixture of hydrocarbons, which are the same as those found on the cuticle. The cuticular hydrocarbon mixture and gland secretion differ, however, in the proportions of components: in the first, linear hydrocarbons are more abundant, while in the second we found a higher quantity of dimethylalkanes. Multivariate statistical analysis has shown that foundresses belonging to different colonies are distinguished on the basis of both the composition of their Dufour gland secretion and cuticular hydrocarbon mixture. Branched hydrocarbons seem to be particularly important in distinguishing females according to their colony. We suggest that the Dufour gland may contribute substances to form the hydrocarbon layer on the cuticle.     

The subepithelial gland in ants: a novel exocrine gland closely associated with the cuticle surface

Two glandular systems were discovered that secrete their products onto the cuticular surface in ants. The first, the subepithelial gland, was previously undescribed in ants, and is found throughout the body just beneath the epithelium. This gland consists of independent secretory units, each made up of a single gland cell and an associated duct cell that penetrates the cuticle. Its ultrastructural appearance is consistent with possible hydrocarbon production. Examining 84 ant species, the subepithelial gland was found in eight subfamilies (out of 13), although not necessarily in all species. In a single ant species, Harpegnathos saltator, it was the epithelium itself that was enlarged and functioned as a gland. The enlarged epithelial cells secrete their products directly onto the cuticle through distinct cuticular crevasses.     

Congruency of hydrocarbon patterns in heterospecific groups of ants: transfer and/or biosynthesis?

Summary In homospecific groups of ants, each species has its own hydrocarbon profile, on the epicuticle and in the postpharyngeal gland (PPG). When reared together in bispecific groups, workers of both species possess each other's hydrocarbons in both locations. The present study investigated two alternative mechanisms by which a mixed odour in artificial groups ofFormica selysi/Manica rubida can be created. Using [1-¹⁴C] sodium acetate as a precursor,de novo biosynthesis of hydrocarbons was demonstrated for both species whether reared in homospecific or mixed-species groups. The newly synthesized hydrocarbons occurred on the epicuticle, internally, and in particularly large amounts in the PPG. As expected from their PPG and epicuticular hydrocarbons composition, workersF. selysi synthesized alkanes and alkenes in comparable amounts irrespective of their rearing scheme. Likewise,M. rubida reared in bispecific groups synthesized mostly alkanes with only negligible amounts of alkenes, according to a ratio characteristic toM. rubida workers from homospecific groups and not toF. selysi workers. During dyadic encounters, a transfer of labeled hydrocarbons between nestmates (conspecific in homospecific groups and allospecific in mixed groups) was observed. These results suggest that the formation of the mixed hydrocarbon profile in artificial groups of ants is the result of a transfer of these chemicals between nestmates rather thande novo biosynthesis of the allospecific hydrocarbons. Behaviours like trophallaxis, grooming and body contact that occurred during the encounters mediated such a transfer.     

Learning and perceptual similarity among cuticular hydrocarbons in ants

Nestmate recognition in ants is based on perceived differences in a multi-component blend of hydrocarbons that are present on the insect cuticle. Although supplementation experiments have shown that some classes of hydrocarbons, such as methyl branched alkanes and alkenes, have a salient role in nestmate recognition, there was basically no information available on how ants detect and perceive these molecules. We used a new conditioning procedure to investigate whether individual carpenter ants could associate a given hydrocarbon (linear or methyl-branched alkane) to sugar reward. We then studied perceptual similarity between a hydrocarbon previously associated with sugar and a novel hydrocarbon. Ants learnt all hydrocarbon-reward associations rapidly and with the same efficiency, regardless of the structure of the molecules. Ants could discriminate among a large number of pairs of hydrocarbons, but also generalised. Generalisation depended both on the structure of the molecule and the animal's experience. For linear alkanes, generalisation was observed when the novel molecule was smaller than the conditioned one. Generalisation between pairs of methyl-alkanes was high, while generalisation between hydrocarbons that differed in the presence or absence of a methyl group was low, suggesting that chain length and functional group might be coded independently by the ant olfactory system. Understanding variations in perception of recognition cues in ants is necessary for the general understanding of the mechanisms involved in social recognition processes based on chemical cues.     

Disentangling the assembly mechanisms of ant cuticular bacterial communities of two Amazonian ant species sharing a common arboreal nest

Bacteria living on the cuticle of ants are generally studied for their protective role against pathogens, especially in the clade of fungus‐growing ants. However, little is known regarding the diversity of cuticular bacteria in other ant host species, as well as the mechanisms leading to the composition of these communities. Here, we used 16S rRNA gene amplicon sequencing to study the influence of host species, species interactions and the pool of bacteria from the environment on the assembly of cuticular bacterial communities on two phylogenetically distant Amazonian ant species that frequently nest together inside the roots system of epiphytic plants, Camponotus femoratus and Crematogaster levior. Our results show that (a) the vast majority of the bacterial community on the cuticle is shared with the nest, suggesting that most bacteria on the cuticle are acquired through environmental acquisition, (b) 5.2% and 2.0% of operational taxonomic units (OTUs) are respectively specific to Ca. femoratus and Cr. levior, probably representing their respective core cuticular bacterial community, and (c) 3.6% of OTUs are shared between the two ant species. Additionally, mass spectrometry metabolomics analysis of metabolites on the cuticle of ants, which excludes the detection of cuticular hydrocarbons produced by the host, were conducted to evaluate correlations among bacterial OTUs and m/z ion mass. Although some positive and negative correlations are found, the cuticular chemical composition was weakly species‐specific, suggesting that cuticular bacterial communities are prominently environmentally acquired. Overall, our results suggest the environment is the dominant source of bacteria found on the cuticle of ants.