Speed and accuracy in nest-mate recognition: a hover wasp prioritizes face recognition over colony odour cues to minimize intrusion by outsiders.

Social insects have evolved sophisticated recognition systems enabling them to accept nest-mates but reject alien conspecifics. In the social wasp, Liostenogaster flavolineata (Stenogastrinae), individuals differ in their cuticular hydrocarbon profiles according to colony membership; each female also possesses a unique (visual) facial pattern. This species represents a unique model to understand how vision and olfaction are integrated and the extent to which wasps prioritize one channel over the other to discriminate aliens and nest-mates. Liostenogaster flavolineata females are able to discriminate between alien and nest-mate females using facial patterns or chemical cues in isolation. However, the two sensory modalities are not equally efficient in the discrimination of 'friend' from 'foe'. Visual cues induce an increased number of erroneous attacks on nest-mates (false alarms), but such attacks are quickly aborted and never result in serious injury. Odour cues, presented in isolation, result in an increased number of misses: erroneous acceptances of outsiders. Interestingly, wasps take the relative efficiencies of the two sensory modalities into account when making rapid decisions about colony membership of an individual: chemical profiles are entirely ignored when the visual and chemical stimuli are presented together. Thus, wasps adopt a strategy to 'err on the safe side' by memorizing individual faces to recognize colony members, and disregarding odour cues to minimize the risk of intrusion from colony outsiders.

High Varroa mite abundance influences chemical profiles of worker bees and mite-host preferences.

Honeybee disappearance is one of the major environmental and economic challenges this century has to face. The ecto-parasitic mite Varroa destructor represents one of the main causes of the worldwide beehive losses. Although halting mite transmission among beehives is of primary importance to save honeybee colonies from further decline, the natural route used by mites to abandon a collapsing colony has not been extensively investigated so far. Here, we explored whether, with increasing mite abundance within the colony, mites change their behaviour to maximize the chances of leaving a highly infested colony. We show that, at low mite abundance, mites remain within the colony and promote their reproduction by riding nurses that they distinguish from foragers by different chemical cuticular signatures. When mite abundance increases, the chemical profile of nurses and foragers tends to overlap, promoting mite departure from exploited colonies by riding pollen foragers.

Ants modulate stridulatory signals depending on the behavioural context.

Insect societies require an effective communication system to coordinate members' activities. Although eusocial species primarily use chemical communication to convey information to conspecifics, there is increasing evidence suggesting that vibroacoustic communication plays a significant role in the behavioural contexts of colony life. In this study, we sought to determine whether stridulation can convey information in ant societies. We tested three main hypotheses using the Mediterranean ant Crematogaster scutellaris: (i) stridulation informs about the emitter'caste; (ii) workers can modulate stridulation based on specific needs, such as communicating the profitability of a food resource, or (iii) behavioural contexts. We recorded the stridulations of individuals from the three castes, restrained on a substrate, and the signals emitted by foragers workers feeding on honey drops of various sizes. Signals emitted by workers and sexuates were quantitatively and qualitatively distinct as was stridulation emitted by workers on different honey drops. Comparing across the experimental setups, we demonstrated that signals emitted in different contexts (restraining vs feeding) differed in emission patterns as well as certain parameters (dominant frequency, amplitude, duration of chirp). Our findings suggest that vibrational signaling represents a flexible communication channel paralleling the well-known chemical communication system.

Hydrocarbon rank signatures correlate with differential oophagy and dominance behaviour in Polistes dominulus foundresses.

Social life offers animals increased fitness opportunities. However, the advantages are not evenly distributed and some individuals benefit more than others. The ultimate advantage of reaching the highest rank in a dominance hierarchy is the achievement of reproduction monopoly. In social insects, dominant individuals and queens keep their reproductive control through differential oophagy of unwanted eggs (egg policing). Egg recognition is the main proximate mechanism for maintaining reproductive dominance. In the social wasp Polistes dominulus, subordinate queens often lay eggs in the presence of the dominant individual. Combining gas chromatography-mass spectrometry (GC-MS) analysis and laboratory bioassays, we found that chemical differences between eggs of subordinate and dominant foundresses can explain the differential success in oophagy enjoyed by dominant individuals. We propose that dominance behaviour is an investigative behaviour as well as a ritualized agonistic behaviour. In fact, the frequency of dominance acts increases with the chemical similarity of the surfaces of dominant- and subordinate-laid eggs. Therefore, dominant individuals probably perform dominance behaviour to test the cuticular signatures of subordinates and so better assess the chemical profiles of subordinate eggs. Finally, we provide evidence that in particular social contexts, subordinate Polistes foundresses can develop ovaries as large as those of dominant individuals but nevertheless lay very few eggs. The subordinates probably lay a limited number of eggs to avoid unnecessary energy loss, as a result of efficient queen policing, but will start laying eggs as soon as the queen fails.

On status badges and quality signals in the paper wasp Polistes dominulus: body size, facial colour patterns and hierarchical rank.

To establish a dominance order, social animals often rely on indicators of fighting to avoid costly aggressive encounters. In some species, individuals use colour patterns to signal their social status. Recent studies claimed that facial markings in the eusocial paper wasp Polistes dominulus are status badges that allow co-foundresses to form a linear hierarchy based on individual quality. Here, we evaluated facial patterns in natural populations of P. dominulus, in its native range, to observe whether the marks reflect overall wasp quality in different contexts. We used the same measures of clypeus patterns used by earlier studies, but did not find that they functioned as status badges. Our analyses showed no evidence that visual markers are related to: (i) size, (ii) probability of surviving winter, (iii) social rank in spring associations, or (iv) health status (assessed by the presence of strepsipteran endoparasites). Size, however, is important. Larger wasps are more likely to survive the winter and to acquire the dominant position in spring associations. Larvae infected with endoparasites become smaller adult wasps. These findings suggest that body size is a reliable quality indicator on which wasps build their social networks, and that clypeus patterning is not involved.

Comparative analysis of epicuticular lipid profiles of sympatric and allopatric field populations of Anopheles gambiae s.s. molecular forms and An. arabiensis from Burkina Faso (West Africa).

We analysed by gas chromatography-mass spectrometry (GC-MS) and Gas Chromatography-Flame Ionization Detector (GC-FID) the epicuticular lipid profiles of field females of the major Afro-tropical malaria vector, Anopheles gambiae. The samples were collected in three villages in Burkina Faso (West Africa), where An. gambiae M and S molecular forms and An. arabiensis live sympatrically. The aim was to compare the cuticular hydrocarbon (CHC) composition of individual field specimens of these three taxa, to highlight possible differences among them. All the samples analysed by GC-MS (55 individuals and eight pools) were characterized by the same 48 CHCs and 10 oxygenated compounds. The 19 most abundant CHCs were quantified in 174 specimens by GC-FID: quantitative intra-taxon differences were found between allopatric populations of both An. arabiensis and S-form. Inter-taxa quantitative differences in the relative abundances of some hydrocarbons between pairs of sympatric taxa were also found, which appear to be mainly linked to local situations, with the possible exception of diMeC(35) between An. arabiensis and S-form. Moreover, MeC(29) shows some degree of differentiation between S- and M-form in all three villages. Possible causes of these differences are discussed.

Social paper wasps as bioindicators: a preliminary research with Polistes dominulus (Hymenoptera Vespidae) as a trace metal accumulator.

The wasps of the genus Polistes (paper wasps), have a worldwide distribution and are widespread in human-built areas. Like other social wasps, they are at the top of food chains and are therefore exposed to the dangers of biomagnification, given that the larvae are fed predominantly with prey that consist of herbivorous insects. The larval faeces, larval fecal masses, in the form of a semi-solid ball, are made up of the residues of the diet of the larva, which are emitted and compressed on the floor of the cell during the larval metamorphosis. Larval fecal masses may accumulate lead (up to 36 times with respect to the adult body), therefore they were used as substrate for the analysis. From the analysis of sample nests of Polistes dominulus in various sites of the urban area of Florence, it emerges that the larval fecal masses are an analytical substrate with which it is possible to distinguish zones with differing degrees of lead pollution. The lead concentration measured in the larval fecal masses turns out to be directly correlated with vehicle traffic density, the main lead source in Florence when the survey was carried out. The notable increase in the lead concentration of larval fecal masses from the rural to the urban nest (11.15 times), in contrast with the much more limited level of pupae (4.39 times), seems to indicate the efficiency of the excretion and/or barrier mechanisms. These wasps seem to be a promising species for biomonitoring lead pollution in order to better understand its dynamics in anthropic ecosystems after the progressive diffusion of unleaded gasoline.

Expression of odorant-binding proteins and chemosensory proteins in some Hymenoptera.

The expression of chemosensory proteins (CSPs) and odorant-binding proteins (OBPs) in individuals of different castes and ages have been monitored in three species of social hymenopterans, Polistes dominulus (Hymenoptera, Vespidae), Vespa crabro (Hymenoptera, Vespidae) and Apis mellifera (Hymenoptera, Apidae), using PCR with specific primers and polyclonal antibodies. In the paper wasp P. dominulus, OBP is equally expressed in antennae, wings and legs of all castes and ages, while CSP is often specifically present in antennae and in some cases also in legs. In the vespine species V. crabro CSP is antennal specific, while OBP is also expressed in legs and wings. The three CSPs and the five OBPs of A. mellifera show a complex pattern of expression, where both classes of proteins include members specifically expressed in antennae and others present in other parts of the body. These data indicate that at least in some hymenopteran species CSPs are specifically expressed in antennae and could perform roles in chemosensory perception so far assigned only to OBPs.

Mating of Xenos vesparum (Rossi) (Strepsiptera, Insecta) revisited.

The controversial mating of the strepsipteran Xenos vesparum was studied to investigate the possible sperm routes for fertilization. The female, which is a neotenic permanent endoparasite of Polistes wasps, extrudes only its anterior region, the "cephalothorax," from the host abdomen. This region has an opening where both mating and larval escape occur. Observations with scanning and transmission electron microscopy revealed spermatozoa not only in the hemocoel, but also in the "ventral canal" (an extragenital duct peculiar to strepsipteran females) and in the "genital ducts" (ectodermal invaginations connecting the ventral canal to the hemocoel) of recently mated females. Xenos vesparum spermatozoa can reach the oocytes either through the hemocoel as a result of a hypodermic insemination, or by moving along the extragenital ducts, which are later used by first instar larvae to escape. The hypothesis of hypodermic insemination is reconsidered in the light of behavioral and ultrastructural evidence.

Recognition of social parasites as nest-mates: adoption of colony-specific host cuticular odours by the paper wasp parasite Polistes sulcifer.

Colonies of the polistine wasp Polistes dominulus are parasitized by the permanent worker-less social parasite Polistes sulcifer. After usurpation of the host colony, parasite females are characterized by a change in the relative proportions of their cuticular hydrocarbons to match those of the host species. In this paper we present evidence from field data and laboratory experiments that P. sulcifer females adopt a colony-specific host odour that facilitates their acceptance by host females of the usurped colony. Presentation experiments demonstrate that parasite females are recognized as foreign individuals by workers of other parasitized nests. We show that the modification of parasite cuticular compounds is sufficient for this recognition. This provides evidence that, after invasion, P. sulcifer queens do not require appeasement or propaganda substances for their acceptance by host colonies. Furthermore, multivariate discriminant analysis of the cuticular hydrocarbon proportions of the parasites after usurpation assigns the parasites together with P. dominulus females of their own host colony. To the authors' knowledge, this is the first confirmation that social parasites adopt colony-specific host odours.

Use of solid-phase microextraction in the investigation of chemical communication in social wasps.

Solid-phase microextraction has been used to investigate chemical communication in several social wasp species. Using the technique to analyse exocrine gland secretions, we demonstrate that the results are comparable with those obtained with the more classical methods that use solvents, eliminating, in many cases, the shortcomings of these methods in insect pheromone analysis. As a result of its simplicity this technique is very suitable for research on the chemical ecology of social wasps, and on insect communication in general.

Ectal mandibular gland in Polistes dominulus (Christ) (Hymenoptera, vespidae): ultrastructural modifications over the secretory cycle.

An ultrastructural study was carried out on the secretory activity of the ectal mandibular gland in the wasp Polistes dominulus (foundress and worker females as well as males). Secretory activity in foundresses proceeds slowly during hibernation and early spring, becoming prominent in late spring and then falling sharply during the summer. This sequential pattern of ultrastructural modifications follows a functional, annual cycle. However, by comparing the subcellular changes in the gland with colonial development, it appears that secretory activity fits in with the specie's social cycle rather than merely following the seasons. The highest levels of secretory activity correspond to the early, critical breeding phases, while activity slows down with an increase in colony protection, based on both primary (passive) and secondary (active) defenses, with the emergence of the workers. These correlations suggest that the ectal mandibular gland secretory product in P. dominulus is involved in chemical nest defense.

Variation in cuticular hydrocarbon signatures, hormonal correlates and establishment of reproductive dominance in a polistine wasp.

In many social insects the relationship between reproductive dominance and physiological correlates is poorly understood. Recent evidence now strongly suggests that cuticular hydrocarbons are important in reproductive differentiation in these societies where they are used as signals of ovarian activity in reproductive females. In this study we investigated the relationship between reproductive dominance, size of the corpora allata (CA, producer of Juvenile Hormone, JH) and the proportions of cuticular hydrocarbons present on the cuticle in overwintering foundresses and both associative (polygynous) and solitary (monogynous) pre-emergence colonies of the social wasp Polistes dominulus. Size of the CA was positively correlated with ovarian development in polygynous colonies. In contrast, solitary foundresses possessed significantly smaller CAs than dominant foundresses from polygynous nests, yet ovarian activity was similar for both female types. CA size variation was associated with variation in cuticular hydrocarbon proportions. Overwintering, solitary, dominant and subordinate (from associative nests) females all possessed distinctive cuticular chemical profiles revealed by multivariate discriminant analyses. Our data indicate that the social environment strongly affects reproductive physiology in this wasp, and we discuss the role of cuticular hydrocarbons in reproductive signaling in P. dominulus and other social insects.

Rank integration in dominance hierarchies of host colonies by the paper wasp social parasite Polistes sulcifer (Hymenoptera, Vespidae).

In multiple-foundress nests of the wasp Polistes dominulus, dominance hierarchies are established among foundresses, and only the dominant (=alpha) individual lays eggs. The alpha female can be distinguished from subordinate females and workers on the basis of the proportions of some hydrocarbons present on the cuticle, suggesting that chemical signaling of her reproductive status could occur. P. dominulus is also the host species of the obligate social parasite Polistes sulcifer. After aggressively usurping host colonies and behaviorally replacing the host alpha female, parasites are characterized by a change in the proportions of their cuticular hydrocarbons to match that of the host cuticular profile at both species and colony levels. In the current study, we demonstrate that P. sulcifer queens also modify their cuticular hydrocarbon proportions after usurpation to match that of the host alpha female. Parasite females, therefore, acquire the dominant rank in host colonies both reproductively and chemically by mimicking the typical alpha profile of the host. Parasite females were not able to fully inhibit ovary development in host foundresses, and 10 days after usurpation, parasites, alpha and beta foundresses show similar chemical profiles and ovarian development.

Fine structure of the Nassonow's gland in the neotenic endoparasitic of female Xenos vesparum (Rossi) (Strepsiptera, Insecta).

Nassonow's gland consists of a number of cells with ducts that open on to the ventral surface of the brood canal in the cephalothoracic region of a neotenic female strepsipteran. The structural organization of the gland is reminiscent of the class 3 of the epidermal gland cells as defined by Noirot and Quennedey [Ann. Rev. Entomol. 19 (1974) 61], which consists of secretory and duct forming cells. The ultrastructure of the Nassonow's gland is described in female Xenos vesparum (Rossi) parasitic in the social wasp Polistes dominulus Christ. The large secretory cells are clustered in groups of three to four, rich in smooth endoplasmic reticulum and produce a secretion made up of lipids. In young females, just before mating, the ultrastructure of the cells and their inclusions indicate that they are active. In old-mated females the Nassonow's gland degenerates. Microvilli line an extracellular cavity and there are pores present in the irregularly thick cuticle of the efferent duct. The small duct forming cells, intermingle with epidermal cells, overlap secretory cells and produce a long efferent duct, the cuticle of which becomes thick close to its opening in the brood canal. Nassonow's gland could be the source of a sex pheromone, which might be capable of attracting the free-living male to a permanently endoparasitic female.

Olfactory cues in host nest detection by the social parasite Polistes sulcifer (Hymenoptera, Vespidae).

Sometimes the nests of the paper wasp Polistes dominulus are parasitized by the obligate social parasite Polistes sulcifer. It is not known how, in the spring, this parasite searches for established nests of its host species. This study investigates the capacity to detect the host nest by olfactory cues alone. In laboratory experiments P. sulcifer females were allowed to choose different options hidden from view: host nest and dummy, various portions of the host nest (larvae, pupae and material), nests or immature brood pertaining to different sympatric species (P. dominulus, P. nimpha and P. gallicus). The parasites proved to be capable of perceiving nest odour and of discriminating between different species of Polistes. The odour of the immature host brood, rather than the nest material, elicits the greatest response in the parasites.

Abdomen stroking behaviour and its possible functions in Polistes dominulus (christ) (hymenoptera, vespidae).

The behaviour of stroking the abdomen over the nest surface has previously been described in females of only a few Polistes wasp species, and was considered to be associated with pheromonal release. This study examines abdomen stroking in solitary and multiple female laboratory colonies of Polistes dominulus during the pre-emergence period. In multiple female colonies stroking behaviour is related to hierarchy: alpha females stroking more than subordinates. However, after removal of the dominant female the subordinate increases her stroking rate. Solitary females also stroke but, unlike alpha females, they do so less during the egg-stage than in subsequent stages of the colonial cycle. It is hypothesized that abdomen stroking cues the immature brood as to which adult female is the dominant individual; this could lead to competition between females on the same nest. This function is also discussed in the context of intra- and inter-specific Polistes parasites, where the behaviour is particularly evident.

Top-down sequencing of Apis dorsata apamin by MALDI-TOF MS and evidence of its inactivity against microorganisms.

Apis mellifera venom is one of the best characterized venoms among Hymenoptera, yet relatively little is known about venom belonging to other species in the genus Apis. Melittin, one of the most important bioactive peptides, has been isolated and characterized in A. mellifera, Apis cerana, Apis dorsata and Apis florea, while apamin has been only characterized in A. mellifera and A. cerana. At present, no information is available about the sequence of A. dorsata apamin. Moreover, while the antiseptic properties of melittin and MCD peptides are well documented, the antimicrobial activity of apamin has never been tested. In the present study, we isolated and characterized apamin from the venom of the giant honeybee A. dorsata. We tested the activity of apamin against bacteria and yeasts in a microbiological assay to gain a more complete understanding of the antimicrobial competence of the medium molecular weight venom fraction. We show that A. dorsata apamin toxin has the same primary sequence as apamin in A. mellifera and A. cerana, yet with a different C-terminal amidation. We did not find any antiseptic activity of apamin against any of the tested microorganisms. We discuss the evolutionary processes connected to the ecological context of venom use that drove the generation of Apis venom complexity.