Nitric oxide radicals are emitted by wasp eggs to kill mold fungi.

Detrimental microbes caused the evolution of a great diversity of antimicrobial defenses in plants and animals. Insects developing underground seem particularly threatened. Here we show that the eggs of a solitary digger wasp, the European beewolf Philanthus triangulum, emit large amounts of gaseous nitric oxide (NO⋅) to protect themselves and their provisions, paralyzed honeybees, against mold fungi. We provide evidence that a NO-synthase (NOS) is involved in the generation of the extraordinary concentrations of nitrogen radicals in brood cells (~1500 ppm NO⋅ and its oxidation product NO2⋅). Sequencing of the beewolf NOS gene revealed no conspicuous differences to related species. However, due to alternative splicing, the NOS-mRNA in beewolf eggs lacks an exon near the regulatory domain. This preventive external application of high doses of NO⋅ by wasp eggs represents an evolutionary key innovation that adds a remarkable novel facet to the array of functions of the important biological effector NO⋅.

Sexual selection and the evolution of male pheromone glands in philanthine wasps (Hymenoptera, Crabronidae).

BACKGROUND: Sexual selection is thought to promote evolutionary changes and diversification. However, the impact of sexual selection in relation to other selective forces is difficult to evaluate. Male digger wasps of the tribe Philanthini (Hymenoptera, Philanthinae) scent mark territories to attract receptive females. Consequently, the organs for production and storage of the marking secretion, the mandibular gland (MG) and the postpharyngeal gland (PPG), are subject to sexual selection. In female Philanthini, these glands are most likely solely subject to natural selection and show very little morphological diversity. According to the hypothesis that sexual selection drives interspecific diversity, we predicted that the MG and PPG show higher interspecific variation in males than in females. Using histological methods, 3D-reconstructions, and multivariate statistical analysis of morphological characters, we conducted a comparative analysis of the MG and the PPG in males of 30 species of Philanthini and three species of the Cercerini and Aphilanthopsini, two related tribes within the Philanthinae. RESULTS: We found substantial interspecific diversity in gland morphology with regard to gland incidence, size, shape and the type of associated secretory cells. Overall there was a phylogenetic trend: Ensuing from the large MGs and small PPGs of male Cercerini and Aphilanthopsini, the size and complexity of the MG was reduced in male Philanthini, while their PPG became considerably enlarged, substantially more complex, and associated with an apparently novel type of secretory cells. In some clades of the Philanthini the MG was even lost and entirely replaced by the PPG. However, several species showed reversals of and exceptions from this trend. Head gland morphology was significantly more diverse among male than among female Philanthinae. CONCLUSION: Our results show considerable variation in male head glands including the loss of an entire gland system and the evolution of a novel kind of secretory cells, confirming the prediction that interspecific diversity in head gland morphology is higher in male than in female Philanthini. We discuss possible causes for the remarkable evolutionary changes in males and we conclude that this high diversity has been caused by sexual selection.

Morphology and ultrastructure of the allomone and sex-pheromone producing mandibular gland of the parasitoid wasp Leptopilina heterotoma (Hymenoptera: Figitidae).

Chemical communication by the parasitoid wasp Leptopilina heterotoma is based largely on (-)-iridomyrmecin. The female wasps use (-)-iridomyrmecin as a defensive allomone, a chemical cue to avoid competition with con- and heterospecific females, and as a major component of their sex pheromone to attract males. Males of L. heterotoma produce (+)-isoiridomyrmecin, which is also used for chemical defense. In this study we show that females and males of L. heterotoma produce the iridomyrmecins in a pair of mandibular glands. Each gland consists of a secretory part composed of class 3 gland cells and their accompanying duct cells, as well as a reservoir bordered by a thin intima. The gland discharges between the mandible base and the clypeus. Males have considerably smaller glands than females, which corresponds to the lower amount of iridomyrmecins produced by males. Chemical analyses of the mandibular gland contents showed that the gland of females contained mainly (-)-iridomyrmecin, as well as low amounts of the other previously described iridoid pheromone compounds, while the glands of males contained only (+)-isoiridomyrmecin. The morphology and sizes of the mandibular glands of males and females of L. heterotoma have evolved to the multi-functional use of iridomyrmecin.

Comparative morphology of the postpharyngeal gland in the Philanthinae (Hymenoptera, Crabronidae) and the evolution of an antimicrobial brood protection mechanism.

BACKGROUND: Hymenoptera that mass-provision their offspring have evolved elaborate antimicrobial strategies to ward off fungal infestation of the highly nutritive larval food. Females of the Afro-European Philanthus triangulum and the South American Trachypus elongatus (Crabronidae, Philanthinae) embalm their prey, paralyzed bees, with a secretion from a complex postpharyngeal gland (PPG). This coating consists of mainly unsaturated hydrocarbons and reduces water accumulation on the prey's surface, thus rendering it unfavorable for fungal growth. Here we (1) investigated whether a North American Philanthus species also employs prey embalming and (2) assessed the occurrence and morphology of a PPG among females of the subfamily Philanthinae in order to elucidate the evolution of prey embalming as an antimicrobial strategy. RESULTS: We provide clear evidence that females of the North American Philanthus gibbosus possess large PPGs and embalm their prey. The comparative analyses of 26 species from six genera of the Philanthinae, using histological methods and 3D-reconstructions, revealed pronounced differences in gland morphology within the subfamily. A formal statistical analysis based on defined characters of the glands confirmed that while all members of the derived tribe Philanthini have large and complex PPGs, species of the two more basal tribes, Cercerini and Aphilanthopsini, possess simple and comparatively small glands. According to an ancestral state reconstruction, the complex PPG most likely evolved in the last common ancestor of the Philanthini, thus representing an autapomorphy of this tribe. CONCLUSION: Prey embalming, as described for P. triangulum and T. elongatus, and now also for P. gibbosus, most probably requires a complex PPG. Hence, the morphology and size of the PPG may allow for inferences about the origin and distribution of the prey embalming behavior within the Philanthinae. Based on our results, we suggest that prey embalming has evolved as an antimicrobial strategy in and is restricted to the tribe Philanthini, which seems to face exceptional threats with regard to fungal infestations of their larval provisions.

Multifaceted defense against antagonistic microbes in developing offspring of the parasitoid wasp Ampulex compressa (Hymenoptera, Ampulicidae).

Effective antimicrobial strategies are essential adaptations of insects to protect themselves, their offspring, and their foods from microbial pathogens and decomposers. Larvae of the emerald cockroach wasp, Ampulex compressa, sanitize their cockroach hosts, Periplaneta americana, with a cocktail of nine antimicrobials comprising mainly (R)-(-)-mellein and micromolide. The blend of these antimicrobials has broad-spectrum antimicrobial activity. Here we explore the spatio-temporal pattern of deployment of antimicrobials during the development from egg to adult as well as their physico-chemical properties to assess how these aspects may contribute to the success of the antimicrobial strategy. Using gas chromatography/mass spectrometry (GC/MS) we show that larvae start sanitizing their food as soon as they have entered their host to feed on its tissue. Subsequently, they impregnate the cockroach carcass with antimicrobials to create a hygienic substrate for cocoon spinning inside the host. Finally, the antimicrobials are incorporated into the cocoon. The antimicrobial profiles on cockroach and wasp cocoon differed markedly. While micromolide persisted on the cockroaches until emergence of the wasps, solid-phase microextraction sampling and GC/MS analysis revealed that (R)-(-)-mellein vaporized from the cockroaches and accumulated in the enclosed nest. In microbial challenge assays (R)-(-)-mellein in the headspace of parasitized cockroaches inhibited growth of entomopathogenic and opportunistic microbes (Serratia marcescens, Aspergillus sydowii, Metarhizium brunneum). We conclude that, in addition to food sanitation, A. compressa larvae enclose themselves in two defensive walls by impregnating the cocoon and the cockroach cuticle with antimicrobials. On top of that, they use vaporous (R)-(-)-mellein to sanitize the nest by fumigation. This multifaceted antimicrobial defense strategy involving the spatially and temporally coordinated deployment of several antimicrobials in solution and vapor form has apparently evolved to reliably protect the larvae themselves and their food against a broad range of antagonistic microbes.

Partner choice and fidelity stabilize coevolution in a Cretaceous-age defensive symbiosis.

Many insects rely on symbiotic microbes for survival, growth, or reproduction. Over evolutionary timescales, the association with intracellular symbionts is stabilized by partner fidelity through strictly vertical symbiont transmission, resulting in congruent host and symbiont phylogenies. However, little is known about how symbioses with extracellular symbionts, representing the majority of insect-associated microorganisms, evolve and remain stable despite opportunities for horizontal exchange and de novo acquisition of symbionts from the environment. Here we demonstrate that host control over symbiont transmission (partner choice) reinforces partner fidelity between solitary wasps and antibiotic-producing bacteria and thereby stabilizes this Cretaceous-age defensive mutualism. Phylogenetic analyses show that three genera of beewolf wasps (Philanthus, Trachypus, and Philanthinus) cultivate a distinct clade of Streptomyces bacteria for protection against pathogenic fungi. The symbionts were acquired from a soil-dwelling ancestor at least 68 million years ago, and vertical transmission via the brood cell and the cocoon surface resulted in host-symbiont codiversification. However, the external mode of transmission also provides opportunities for horizontal transfer, and beewolf species have indeed exchanged symbiont strains, possibly through predation or nest reuse. Experimental infection with nonnative bacteria reveals that--despite successful colonization of the antennal gland reservoirs--transmission to the cocoon is selectively blocked. Thus, partner choice can play an important role even in predominantly vertically transmitted symbioses by stabilizing the cooperative association over evolutionary timescales.

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.

Double deception: ant-mimicking spiders elude both visually- and chemically-oriented predators.

Biological mimicry is often multimodal, in that a mimic reinforces its resemblance to another organism via different kinds of signals that can be perceived by a specific target audience. In this paper we describe a novel scenario, in which a mimic deceives at least two distinct audiences, each of which relies primarily on a different sensory modality for decision-making. We have previously shown that Peckhamia picata, a myrmecomorphic spider that morphologically and behaviorally resembles the ant Camponotus nearcticus, experiences reduced predation by visually-oriented jumping spiders. Here we report that Peckhamia also faces reduced aggression from spider-hunting sphecid wasps as well as from its model ant, both of which use chemical cues to identify prey. We also report that Peckhamia does not chemically resemble its model ants, and that its total cuticular hydrocarbons are significantly lower than those of the ants and non-mimic spiders. Although further studies are needed to clarify the basis of Peckhamia's chemically-mediated protection, to our knowledge, such 'double deception,' in which a single organism sends misleading visual cues to one set of predators while chemically misleading another set, has not been reported; however, it is likely to be common among what have until now been considered purely visual mimics.

Larvae of the parasitoid wasp Ampulex compressa sanitize their host, the American cockroach, with a blend of antimicrobials.

Food resources contaminated with spoilage or pathogenic microorganisms pose severe problems to all higher organisms. Here, we describe a food-hygienic strategy of the emerald cockroach wasp Ampulex compressa. The wasp larvae develop on and inside the American cockroach Periplaneta americana, a host that can harbor various putrefactive microbes, as well as human and insect pathogens. From P. americana, we isolated the Gram-negative bacterium Serratia marcescens, which is a potent entomopathogen that can rapidly kill insect larvae. It is also known as a food contaminant and as an opportunistic human pathogen. Using behavioral observations and chemical analyses, we demonstrated that A. compressa larvae impregnate their cockroach hosts from inside with large amounts of an oral secretion containing a blend of γ-lactones and isocoumarins with (R)-(-)-mellein [(R)-(-)-3,4-diydro-8-hydroxy-3-methylisocoumarin] and micromolide [(4R,9Z)-octadec-9-en-4-olide] as dominant components. We fractionated hexane extracts of the secretion and investigated the antimicrobial properties of the fraction containing the lactones and isocoumarins, as well as of synthetic (R)-(-)-mellein and micromolide, against S. marcescens and a Gram-positive bacterium, Staphylococcus hyicus, in broth microdilution assays. The test fraction inhibited growth of both tested bacteria. The activity of the fraction against S. marcescens was explained by (R)-(-)-mellein alone, and the activity against S. hyicus was explained by the combined action of (R)-(-)-mellein and micromolide. Our data suggest that the specific combination of antimicrobials in the larval secretion provides an effective frontline defense against the unpredictable spectrum of microbes that A. compressa larvae may encounter during their development inside their cockroach hosts.

Refining the roots of the beewolf-Streptomyces symbiosis: antennal symbionts in the rare genus Philanthinus (Hymenoptera, Crabronidae).

Insects engage in symbiotic associations with a large diversity of beneficial microorganisms. While the majority of well-studied symbioses have a nutritional basis, several cases are known in which bacteria protect their host from pathogen infestation. Solitary wasps of the genera Philanthus and Trachypus (beewolves; Hymenoptera, Crabronidae) cultivate the actinomycete "Candidatus Streptomyces philanthi" in specialized antennal gland reservoirs. The symbionts are transferred to the larval cocoon, where they provide protection against pathogenic fungi by producing at least nine different antibiotics. Here we investigated the closest relatives of Philanthus and Trachypus, the rare genus Philanthinus, for the presence of antennal gland reservoirs and symbiotic streptomycetes. Molecular analyses identified "Ca. Streptomyces philanthi" in reservoirs of Philanthinus quattuordecimpunctatus. Phylogenies based on the 16S rRNA gene suggest that P. quattuordecimpunctatus may have acquired "Ca. Streptomyces philanthi" by horizontal transfer from other beewolf species. In histological sections and three-dimensional reconstructions, the antennal gland reservoirs were found to occupy six antennal segments (as opposed to only five in Philanthus and Trachypus) and to be structurally less complex than those of the evolutionarily more derived genera of beewolves. The presence of "Ca. Streptomyces philanthi" in antennal glands of Philanthinus indicates that the symbiosis between beewolves and Streptomyces bacteria is much older than previously thought. It probably evolved along the branch leading to the monophyletic tribe Philanthini, as it seems to be confined to the genera Philanthus, Trachypus, and Philanthinus, which together comprise 172 described species of solitary wasps.

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.

The chemistry of the postpharyngeal gland of female European beewolves.

Females of the European beewolf, Philanthus triangulum, possess a large glove-shaped gland in the head, the postpharyngeal gland (PPG). They apply the content of the PPG to their prey, paralyzed honeybees, where it delays fungal infestation. Here, we describe the chemical composition of the gland by using combined GC-MS, GC-FTIR, and derivatization. The PPG of beewolves contains mainly long-chain unsaturated hydrocarbons (C23-C33), lower amounts of saturated hydrocarbons (C14-C33), and minor amounts of methyl-branched hydrocarbons (C17-C31). Additionally, the hexane-soluble gland content is comprised of small amounts of an unsaturated C25 alcohol, an unknown sesquiterpene, an octadecenylmethylester, and several long-chain saturated (C25, C27) and unsaturated (C23-C27) ketones, some of which have not yet been reported as natural products. Surprisingly, we found a dimorphism with regard to the major component of the PPG with some females having (Z)-9-pentacosene, whereas others have (Z)-9-heptacosene as their predominant component. The biological relevance of the compounds for the prevention of fungal growth on the prey and the significance of the chemical dimorphism are discussed.

Chemical profiles of mated and virgin queens, egg-laying intermorphs and workers of the ant Crematogaster smithi.

Many colonies of the North American ant Crematogaster smithi contain a "third female caste" in addition to queens and workers. These "intermorphs" are morphological intermediate of queens and workers and have well-developed ovaries but lack a spermatheca for the storage of sperm. They are specialised for laying large numbers of unfertilised, viable eggs, most of which serve as food for larvae and adults, though a few may eventually develop into males. Based on the assumption that cuticular hydrocarbons (CHCs) in social insects honestly signal the reproductive status of an individual we investigated the CHC of mated mature queens, virgin queens, intermorphs and workers. We expected intermorphs to show chemical profiles intermediate between those of mated queens and non-reproductive workers. A discriminant analysis of the chemical profiles reliably separated queens, virgin queens, and workers, but failed to distinguish between queens and intermorphs even though workers were apparently capable of doing so.

A cuckoo in wolves' clothing? Chemical mimicry in a specialized cuckoo wasp of the European beewolf (Hymenoptera, Chrysididae and Crabronidae).

BACKGROUND: Host-parasite interactions are among the most important biotic relationships. Host species should evolve mechanisms to detect their enemies and employ appropriate counterstrategies. Parasites, in turn, should evolve mechanisms to evade detection and thus maximize their success. Females of the European beewolf (Philanthus triangulum, Hymenoptera, Crabronidae) hunt exclusively honeybee workers as food for their progeny. The brood cells containing the paralyzed bees are severely threatened by a highly specialized cuckoo wasp (Hedychrum rutilans, Hymenoptera, Chrysididae). Female cuckoo wasps enter beewolf nests to oviposit on paralyzed bees that are temporarily couched in the nest burrow. The cuckoo wasp larva kills the beewolf larva and feeds on it and the bees. Here, we investigated whether H. rutilans evades detection by its host. Since chemical senses are most important in the dark nest, we hypothesized that the cuckoo wasp might employ chemical camouflage. RESULTS: Field observations suggest that cuckoo wasps are attacked by beewolves in front of their nest, most probably after being recognized visually. In contrast, beewolves seem not to detect signs of the presence of these parasitoids neither when these had visited the nest nor when directly encountered in the dark nest burrow.In a recognition bioassay in observation cages, beewolf females responded significantly less frequently to filter paper discs treated with a cuticular extract from H. rutilans females, than to filter paper discs treated with an extract from another cuckoo wasp species (Chrysis viridula). The behavior to paper discs treated with a cuticular extract from H. rutilans females did not differ significantly from the behavior towards filter paper discs treated with the solvent only.We hypothesized that cuckoo wasps either mimic the chemistry of their beewolf host or their host's prey. We tested this hypothesis using GC-MS analyses of the cuticles of male and female beewolves, cuckoo wasps, and honeybee workers. Cuticle extracts of Hedychrum nobile (Hymenoptera: Chrysididae) and Cerceris arenaria (Hymenoptera: Crabronidae) were used as outgroups. There was little congruence with regard to cuticular compounds between H. rutilans females and honeybees as well as females of C. arenaria and H. nobile. However, there was a considerable similarity between beewolf females and H. rutilans females. Beewolf females show a striking dimorphism regarding their cuticular hydrocarbons with one morph having (Z)-9-C25:1 and the other morph having (Z)-9-C27:1 as the major component. H. rutilans females were more similar to the morph having (Z)-9-C27:1 as the main component. CONCLUSION: We conclude that H. rutilans females closely mimic the composition of cuticular compounds of their host species P. triangulum. The occurrence of isomeric forms of certain compounds on the cuticles of the cuckoo wasps but their absence on beewolf females suggests that cuckoo wasps synthesize the cuticular compounds rather than sequester them from their host. Thus, the behavioral data and the chemical analysis provide evidence that a specialized cuckoo wasp exhibits chemical mimicry of the odor of its host. This probably allows the cuckoo wasp to enter the nest with a reduced risk of being detected by olfaction and without leaving traitorous chemical traces.

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.

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.

Males of a solitary wasp possess a postpharyngeal gland.

The postpharyngeal gland has long been thought to occur only in ants. Here we characterize, by use of light and electron microscopy as well as 3D reconstruction based on nuclear magnetic resonance (NMR) imaging data, a large cephalic gland reservoir of males of a solitary digger wasp, the European beewolf, Philanthus triangulum. Several lines of evidence suggest that this reservoir is a postpharyngeal gland. The gland reservoir originates from the posterior part of the pharynx and consists of two pairs of unbranched tubular structures that occupy a large portion of the head capsule. Its wall is composed of a unicellular epithelium that is lined by a cuticle. The gland contains a blend of hydrocarbons and compounds with functional groups, and we show that the hydrocarbon fraction of the pheromone is congruent with the hydrocarbons on the cuticle. We discuss the implications of our findings for the evolution of the postpharyngeal gland in ants.

Food wrapping with the postpharyngeal gland secretion by females of the European beewolf Philanthus triangulum.

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.

A selfish function of a "social" gland? A postpharyngeal gland functions as a sex pheromone reservoir in males of the solitary wasp Philanthus triangulum.

The postpharyngeal gland (PPG) has long been assumed to be restricted to ants, where it mainly functions in the maintenance of social integrity. Recently, a PPG has been described in both sexes of a solitary digger wasp, the European beewolf, Philanthus triangulum (Hymenoptera, Crabronidae). Female beewolves use the contents of their PPG to embalm their honeybee prey to delay microbial growth. Here we show that in male beewolves, the PPG serves as a reservoir of the pheromone used to scent-mark their territories. Gas chromatography-mass spectrometry (GC-MS) analysis of PPG contents identified 55 substances including long-chain aliphatic hydrocarbons, and 13 substances with functional groups. The composition was consistent with the composition of the marking pheromone of male European beewolves described earlier. Comparisons of the PPG contents, and total-head extracts showed a strong congruency, suggesting that total-head extracts can be used for the analysis of marking secretion in beewolves. Furthermore, we found a dimorphism in the composition of the PPG contents, based on significant differences in the proportions of seven compounds between the two morphs.