Use of Dufour's gland secretion in nest defence and brood nutrition by hover wasps (Hymenoptera, Stenogastrinae)

Social wasps of the subfamily Stenogastrinae produce an abdominal secretion that is used in two distinct biological contexts. First, the secretion plays an important role in larval nutrition where it serves as a substrate in which food is placed by the adults for eventual consumption by the larvae. Second, in several species, females apply the same secretion to the substrate on which their nests are constructed, where it constitutes a sticky barrier that defends the immature brood from predation by ants. This paper describes for the first time ant guard construction behaviour of three species of stenogastrine wasps belonging to the genera Eustenogaster and Liostenogaster. The identification of compounds making up these secretions was also performed by gas chromatography-mass spectrometry. Ant guards and brood secretions were similar, with saturated and unsaturated long chain hydrocarbons and alcohols as major components. We further confirm that the glandular source of abdominal secretion is the Dufour's gland. This gland contains the same hydrocarbons, and in the same proportions as ant guards and brood secretion. We discuss the fundamental importance of Dufour's gland secretion in the social life of these wasps by comparing species with and without ant guards within the subfamily.     

Post-pollination emission of a repellent compound in a sexually deceptive orchid: a new mechanism for maximising reproductive success?

The flowers of the sexually deceptive orchid Ophrys sphegodes are pollinated by pseudocopulating males of the solitary bee Andrena nigroaenea. We investigated the changes in odor emission and reduced attractiveness that occur after pollination in these plants. We analyzed floral odor of unpollinated and pollinated flowers by gas chromatography and compared relative and absolute amounts of electrophysiologically active compounds. Headspace odor samples of O. sphegodes flowers showed a significant increase in absolute and relative amounts of all-trans-farnesyl hexanoate after pollination. Flower extracts also indicated an increase of farnesyl hexanoate after pollination. The total amount of the other physiologically active odor compounds decreased slightly. Farnesyl hexanoate is a major constituent of the Dufour's gland secretion in females of the pollinator bees, A. nigroaenea, where it functions in the lining of the brood cells. Furthermore, this compound lowers the number of copulation attempts in males. In dual-choice tests, we showed that flowers artificially scented with an amount of farnesyl hexanoate equal to the increased amount after pollination were significantly less attractive than flowers treated with solvent only. We propose that the increased production of farnesyl hexanoate in pollinated flowers is a signal to guide pollinators to unpollinated flowers of the inflorescence, which represents a new mechanism in this pollination system.     

Behavioral responses to the alarm pheromone of the ant Camponotus obscuripes (Hymenoptera: Formicidae)

The alarm pheromone of the ant Camponotus obscuripes (Formicinae) was identified and quantified by gas chromatography (GC) and gas chromatography-mass spectrometry (GC-MS). Comparisons between alarm pheromone components and extracts from the major exocrine gland of this ant species revealed that the sources of its alarm pheromone are Dufour's gland and the poison gland. Most components of Dufour's gland were saturated hydrocarbons. n-Undecane comprised more than 90% of all components and in a single Dufour's gland amounted to 19 microg. n-Decane and n-pentadecane were also included in the Dufour's gland secretion. Only formic acid was detected in the poison gland, in amounts ranging from 0.049 to 0.91 microl. This ant species releases a mixture of these substances, each of which has a different volatility and function. When the ants sensed formic acid, they eluded the source of the odor; however, they aggressively approached odors of n-undecane and n-decane, which are highly volatile. In contrast, n-pentadecane, which has the lowest volatility among the identified compounds, was shown to calm the ants. The volatilities of the alarm pheromone components were closely related to their roles in alarm communication. Highly volatile components vaporized rapidly and spread widely, and induced drastic reactions among the ants. As these components became diluted, the less volatile components calmed the excited ants. How the worker ants utilize this alarm communication system for efficient deployment of their nestmates in colony defense is also discussed herein.     

Queen-signal modulation of worker pheromonal composition in honeybees

Worker sterility in honeybees is neither absolute nor irreversible. Whether under queen or worker control, it is likely to be mediated by pheromones. Queen-specific pheromones are not exclusive to queens; workers with activated ovaries also produce them. The association between ovarian activation and queen-like pheromone occurrence suggests the latter as providing a reliable signal of reproductive ability. In this study we investigated the effect of queen pheromones on ovary development and occurrence of queen-like esters in workers' Dufour's gland. Workers separated from the queenright compartment by a double mesh behaved like queenless workers, activating their ovaries and expressing a queen-like Dufour's gland secretion, confirming that the pheromones regulating both systems are non-volatile. Workers with developed ovaries produced significantly more secretion than sterile workers, which we attribute primarily to increased ester production. Workers separated from the queenright compartment by a single mesh displayed a delayed ovarian development, which we attribute to interrupted transfer of the non-volatile pheromone between compartments. We suggest that worker expression of queen-like characters reflects a queen-worker arms race; and that Dufour's gland secretion may provide a reliable signal for ovarian activation. The associative nature between ovary development and Dufour's gland ester production remains elusive.     

Host-parasite relationships in six species ofSphecodes bees and their halictid hosts: Nest intrusion,intranidal behavior,and Dufour's gland volatiles (Hymenoptera: Halictidae)

Nest invasion behavior was studied in six kleptoparasiticSphecodes species at four nesting sites of their respective social and solitary hosts.Sphecodes females preferred to enter unguarded nests. Nest intruding strategies observed in the differentSphecodes species did not depend on whether host species were solitary or social, as long as the nesting cycle of a social host was in the solitary stage (i.e., a single host female). Observation of intranidal behavior revealed thatSphecodes monilicornis females kill all host individuals within an usurped nest. They stay in the nest for several hours, laying eggs in adequately provisioned brood cells. Gas chromatography-mass spectrometry analyses of Dufour's gland secretions revealed species-specific compositions. Qualitative comparisons of whole patterns and quantitative comparisons considering the predominant hydrocarbons common to both host and parasite contradict the hypothesis of chemical mimetism, a mechanism supposed to permit parasite intrusion by qualitatively similar odor bouquets in host and parasite females.     

Source of the host marking pheromone in the egg parasitoid Trissolcus basalis (Hymenoptera: Scelionidae)

After oviposition, Trissolcus basalis females always mark the host's surface, depositing host marking substances for herself and to warn other ovipositing females. The perception of these host marking substances, probably through the antennae, can induce the female to leave and seek healthy hosts. Parasitoid females exposed to conspecific parasitized egg masses left the host egg masses significantly more often than when exposed to non-parasitized egg masses. More egg mass leaving behavior also was observed when the egg masses were treated with Dufour's gland secretion but not when treated with secretion from the common oviducts. The common oviduct has a secretory epithelium that produces electron-dense vesicles, probably containing proteinaceous substances. The secretory cells of the accessory gland, Dufour's gland, contain electron-lucid vesicles, whose secretion appears to be a lipid similarly to that found in pheromone secreting glands. Ultrastructural and behavioral evidence suggests that Dufour's gland is the host marking pheromone source.     

The influence of brood comb cell size on the reproductive behavior of the ectoparasitic mite Varroa destructor in Africanized honey bee colonies

Africanized honey bees (Apis mellifera, Hymenoptera: Apidae) in Brazil are tolerant of infestations with the exotic ectoparasitic mite, Varroa destructor (Mesostigmata: Varroidae), while the European honey bees used in apiculture throughout most of the world are severely affected. Africanized honey bees are normally kept in hives with both naturally built small width brood cells and with brood cells made from European-sized foundation, yet we know that comb cell size has an effect on varroa reproductive behavior. Three types (sizes) of brood combs were placed in each of six Africanized honey bee colonies: new (self-built) Africanized comb, new Italian comb (that the bees made from Italian-sized commercial foundation), and new Carniolan comb (built naturally by Carniolan bees). About 100 cells of each type were analyzed in each colony. The Africanized comb cells were significantly smaller in (inner) width (4.84 mm) than the European-sized comb cells (5.16 and 5.27 mm for Italian and Carniolan cells, respectively). The brood cell infestation rates (percentage cells infested) were significantly higher in the Carniolan-sized comb cells (19.3%) than in the Italian and Africanized cells (13.9 and 10.3%, respectively). The Carniolan-sized cells also had a significantly larger number of invading adult female mites per 100 brood cells (24.4) than did the Italian-sized cells (17.7) and the natural-sized Africanized worker brood cells (15.6). European-sized worker brood cells were always more infested than the Africanized worker brood cells in the same colony. There was a highly significant correlation (P<0.01) between cell width and the rate of infestation with varroa in four of the six colonies. The small width comb cells produced by Africanized honey bees may have a role in the ability of these bees to tolerate infestations by Varroa destructor, furthermore it appears that natural-sized comb cells are superior to over-sized comb cells for disease resistance.     

Social Insect Pheromones: Their Chemistry and Function

Exocrine secretions of social insects are often characterizedby extraordinarily complex mixtures of natural products. Thus,chemical communication in social insects must be interpretedin terms of signals generated by multicomponent systems, theindividual constituents of which can affect the informationalcontent of the message. Alarm pheromones have been identified chiefly in three subfamiliesof ants and their distribution appears to be chemosystematicallysignificant. Myrmicine genera emphasize 3-alkanones as alarmreleasers, whereas methyl ketones, primarily of terpenoidalorigin, are widely utilized as alarm pheromones in the subfamilyDolichoderinae. Formicine species may employ formic acidas analarm pheromone in addition to the compounds produced in themandibular and Dufour's glands. The mandibular gland pheromonesare chiefly acyclic monoterpene aldehydes (e.g., citronellal)which are relatively low boiling compounds. Higher boiling n-alkanesare produced in the Dufour's glands and may serve as more persistentreleasers of alarm behavior. Alarm pheromones as well as thecaste-specific pheromones of male bees and ants, probably alsoserve as defensive products. In many cases it is likely thatpheromones were originally utilized as defensive compounds andtheir communicative function is a secondary development.     

Brood comb construction by the stingless bees Tetragonula hockingsi and Tetragonula carbonaria

Tetragonula hockingsi and T. carbonaria are two closely related species of Australian stingless bees. The primary species-specific character is the architecture of the brood comb. The brood comb of T. hockingsi is an open lattice comprising clumps of about ten cells that are connected by vertical pillars. In contrast, in T. carbonaria the brood comb is a compact spiral in which all brood cells (except on the margins) are connected by their walls to adjacent cells at the same height. We made detailed observations of the cell construction process in two colonies of each species. From these observations we formed a species-specific hypothesis about the algorithm followed by the bees during cell construction. The two algorithms allowed us to make predictions about the locations of new cells. Both T. hockingsi and T. carbonaria share a preference for constructing new brood cells in the clefts formed by two or three adjacent existing brood cells, but there are differences in detail for other components of the building process. The fundamental difference in the cell construction process of the two species is that for T. hockingsi, when a cluster of cells contains ten cells, the next cell added to the cluster is offset upwards by half a cell length, or, less often, a vertical pillar rather than a new cell is constructed. In T. carbonaria, cell construction is continuous at the comb margin so that there are no gaps between cells. Furthermore, it seems that T. hockingsi only makes use of local knowledge of the brood comb when deciding to place new brood cells, whereas T. carbonaria could make some building decisions based on knowledge of the total structure. We translated the species-specific algorithms into agent-based lattice swarm computer simulations of the cell construction process for the two species. These simulations produced representations of brood combs that are similar to those seen in vivo, suggesting that our biological rules are realistic.     

Multitrophic effects of experimental changes in plant diversity on cavity-nesting bees, wasps, and their parasitoids

Plant diversity changes can impact the abundance, diversity, and functioning of species at higher trophic levels. We used an experimental gradient in grassland plant diversity ranging from 1 to 16 plant species to study multitrophic interactions among plants, cavity-nesting bees and wasps, and their natural enemies, and analysed brood cell density, insect diversity (species richness), and bee and wasp community similarity over two consecutive years. The bee and wasp communities were more similar among the high (16 species) diversity plots than among plots of the lower diversity levels (up to 8 species), and a more similar community of bees and wasps resulted in a more similar community of their parasitoids. Plant diversity, which was closely related to flower diversity, positively and indirectly affected bee diversity and the diversity of their parasitoids via increasing brood cell density of bees. Increasing plant diversity directly led to higher wasp diversity. Parasitism rates of bees and wasps (hosts) were not affected by plant diversity, but increased with the diversity of their respective parasitoids. Decreases in parasitism rates of bees arose from increasing brood cell density of bees (hosts), whereas decreasing parasitism rates of wasps arose from increasing wasp diversity (hosts). In conclusion, decreases in plant diversity propagated through different trophic levels: from plants to insect hosts to their parasitoids, decreasing density and diversity. The positive relationship between plant diversity and the community similarity of higher trophic levels indicates a community-stabilising effect of high plant diversity.     

Fine structure of the accessory glands of the female genital tract of the ichneumonid Pimpla turionellae (Insecta,Hymenoptera)

Summary The female accessory glands include the tubular poison gland, the paired, lemon-shaped uterus glands, and Dufour's gland, an unbranched tubular organ. They consist essentially of a single layer of epithelium cells surrounded by a basement membrane. The lumen is lined by cuticle. The proteinaceous secretion of the poison gland is released into intracellular ducts provided with microvilli, each connected to a channel lined with cuticle which leads to the central lumen of the gland. The channel is formed by special canal cells. Nerve endings are interspersed among the gland cells. The uterus gland consists of four cell types derived from a single type of precursor cell found in newly hatched wasps. Type I cells are covered by type II cells and are thus without contact to the luminal surface of the gland. They contain stacks or whorls of mitochondria and smooth cisternae in an alternating arrangement. Vesicles with a secretory product are found in cells of types II and III. Deep anastomosing infoldings of the plasmalemma, stabilized by microtubules and dense material at the branchings, are characteristic for type II cells. Most secretory vesicles are found in type III cells, the prevalent cell type which is thought to be the source of the lipoprotein secretion. Coated vesicles are present at deep infoldings of the plasmalemma. The greatly enlarged apical surface area of type IV cells and the presence of mitochondria in slender outgrowths is suggestive of an osmoregulatory function. In Dufour's gland, two cell types appear in succession, the first with a very dense cytoplasm, the second with dense inclusions and many seemingly empty vesicles of smooth endoplasmic reticulum. The secretion products, lecithin and a cholesterol ester, are thought to be formed by the second cell type. The dense inclusion might be lecithin, which reacts with osmium tetroxide. The cholesterol ester could have been washed out of the empty vesicles by the embedding procedure.     

The orientation inducer pheromone of the fire ant Solenopsis invicta

Abstract Foraging ants recruit nestmate workers to food sources by a variety of mechanisms. We report that one behavioural subcategory of the recruitment pheromone complex of Solenopsis invicta Buren involves orientation induction. The orientation inducer pheromone exerts its effects by changing the physiological state of the recipient rather than by releasing a measurable behaviour. Some ant species use a physical 'waggle' behaviour to motivate (change physiological state) nestmate workers to follow their chemical trail. The orientation inducer pheromone can be interpreted as a chemical analogue of the physical 'waggle' inducing effects. This behaviour is not elicited by the recruitment pheromone components responsible for orientation and/or attraction. Each of these behavioural categories is mediated by a different blend of chemicals from the Dufour's gland. Activity-concentration thresholds indicate that the attraction and inducer part of the recruitment pheromone require about 250 times more worker equivalents for a response than the orientation pheromone. Therefore, the recruitment sub-categories are differentially activated by the amount of Dufour's gland material released.     

Functional flexibility of the honey bee hypopharyngeal gland in a dequeened colony

The role of the worker honey bee Apis mellifera L. changes depending on age after eclosion (age polyethism): young workers (nurse bees) take care of their brood by synthesizing and secreting brood food (royal jelly), while older workers (foragers) forage for nectar and process it into honey. Previously, we showed that the major proteins synthesized in the hypopharyngeal gland of the worker change from brood food proteins to alpha-glucosidase at the single secretory cell level in parallel with this age polyethism [Kubo et al., J. Biochem. 119, 291-295 (1996); Ohashi et al., Eur. J. Biochem. 249, 797-802 (1997)]. Here, we examined whether the function of the hypopharyngeal gland has flexibility depending on the colony conditions, by creating a dequeened colony in which the older workers were compelled to feed the drone larvae. It was found that most of the older workers in the dequeened colony synthesized brood food proteins as did nurse bees. Furthermore, the percentage of workers that synthesized brood food proteins was maintained at 80-90% of the total workers for at least two months, as in a normal colony. These results indicate that the function of the hypopharyngeal gland cells of the worker has flexibility and can, if necessary, be maintained as that of the nurse bee, depending on the condition of the colony.     

Females of the European beewolf preserve their honeybee prey against competing fungi

1. Females of the European beewolf Philanthus triangulum (Hymenoptera, Sphecidae) provision brood cells with paralysed honeybees as larval food. Because brood cells are located in warm, humid locations there is a high risk of microbial decomposition of the provisions. Low incidence of fungus infestation (Aspergillus sp.) in nests in the field suggested the presence of an anti‐fungal adaptation. 2. To test whether the paralysis caused the protection from fungus infestation, the timing of fungus growth on bees that were freeze‐killed, paralysed but not provisioned, and provisioned regularly by beewolf females was determined. Fungus growth was first detected on freeze‐killed bees, followed by paralysed but not provisioned bees. By contrast, fungus growth on provisioned bees was delayed greatly or even absent. Thus, paralysis alone is much less efficient in delaying fungus growth than is regular provisioning. 3. Observations of beewolves in their nests revealed that females lick the body surface of their prey very thoroughly during the period of excavation of the brood cell. 4. To separate the effect of a possible anti‐fungal property of the brood cell and the licking of the bees, a second experiment was conducted. Timing of fungus growth on paralysed bees did not differ between artificial and original brood cells. By contrast, fungus growth on bees that had been provisioned by a female but were transferred to artificial brood cells was delayed significantly. Thus, the treatment of the bees by the female wasp but not the brood cell caused the delay in fungus growth. 5. Beewolf females most probably apply anti‐fungal chemicals to the cuticle of their prey. This is the first demonstration of the mechanism involved in the preservation of provisions in a hunting wasp. Some kind of preservation of prey as a component of parental care is probably widespread among hunting wasps and might have been a prerequisite for the evolution of mass provisioning.     

Honeybee egg-laying workers mimic a queen signal

Summary. In the honeybee, Dufour's gland secretion is caste specific and constitutes a component of the multi-sourced queen signal. As predicted, it is attractive to workers, which form a retinue around the scented source. Bioassays reveal the ester fraction and not the hydrocarbons to be the active constituents. This function of the esters was corroborated by assays with the synthetic queen-esters mixture, which successfully mimicked the queen's secretion. As predicted from the queen-like secretion exhibited by egg-laying workers, their glandular secretion was also attractive to nestmates, albeit to a lesser degree than that of the queen; while that of non-egg-laying workers was totally inactive. The evolution of the multiple queen signals in honeybees can be regarded as a component in an arms race between queen and workers. We hypothesize that in response to a reduced sensitivity to a certain queen signal, queen honeybees were selected to develop an alternative signaling-source. Dufour's gland seems to be one of these sources.     

Subsociality in halictine bees

To look for the occurrence and the significance of brood care in social evolution, I reared six eusocial halictine bee species in laboratory cages enabling the observation of intranest behaviour: Lasioglossum (Evylaeus) laticeps, L. (E.) pauxillum, L. (E.) nigripes, L. (E.) euboeensis, Halictus (Halictus) scabiosae and L. (E.) fulvicorne. All of them were subsocial, each mother caring for her brood. Brood cells were sealed after oviposition with earthen plugs; they were then reopened, visited and closed again. These observations plus the reports in the literature on eleven eusocial species indicate that seventeen species of eusocial halictine bees provide parental care, i.e. are subsocial. Brood care, subsociality, is strongly associated with eusociality. To study reversal from eusociality to subsociality, I have reared the non-eusocial form of two species within which there are or have been eusocial forms: Halictus (H.) rubicundus and Lasioglossum (E.) fratellum. They are secondarily solitary, having lost worker brood. However, both species still show brood care. This suggests that in transitions to eusociality, brood care antedated eusociality. To further examine this issue I reared two truly solitary species that are not derived from eusocial ancestors: Lasioglossum (E.) villosulum and L. (L.) quadrinotatum. Unlike secondarily solitary species, females of both these species close their brood cells after oviposition and ignore their progeny thereafter. This association strongly suggests that the subsocial route with maternal brood care is the route to eusociality in halictine bees.     

Biology and social behaviour of three species ofAnischnogaster (Vespidae,Stenogastrinae) in Papua New Guinea

Summary Level of social organization, adult behaviour, size and development of the brood in three species ofAnischnogaster are described. The normal colony size in all the three species was one female per nest, and colony size never exceeded two females per nest. The social interactions, which were only observed in one species, did not include any marked dominance behaviour, but there was clear caste differentiation, with the older female guarding the nest while the younger female foraged for food. InAnischnogaster sp. A only some eggs and larvae have abdominal secretion, while no secretion at all was found on the eggs and larvae ofA. laticeps. In spite of this, the Dufour's gland was found to be well developed. The significance of this is discussed. Females ofA. laticeps were found to fall into two groups distinguished by the length of the sting. The larvae seem quite similar to those of other Stenogastrinae and have, apparently, only four instars.     

Ethology of Hygienic Behaviour in the Honey Bee Apis mellifera L. (Hymenoptera: Apidae): Behavioural repertoire of Hygienic bees

Hygienic behaviour performed by middle‐aged worker bees is an important intranidal task in colonies of the honey bee Apis mellifera (L.). It comprises detecting diseased brood in the larval and pupal stages and removing all such infected brood, thereby decreasing the incidence of infection. Hygienic behaviour consists of two task‐components: uncapping cells and removing the cell contents. The aim of this study was to observe bees performing hygienic behaviour to determine their age at performance of the behaviour and to describe their behavioural repertoire. The bees performing hygienic behaviour were middle‐aged bees, younger than foragers. In the colonies where the behaviours of individual bees were observed, all bees performing the hygienic behaviour were seen to exhibit both the components, though at different frequencies. One behavioural class performed the task of uncapping cells at higher frequencies than the task of removing cell contents, while another class performed both tasks to the same extent. While these two classes had higher frequencies of the tasks comprising the hygienic behaviour but lower frequencies of other common behaviours in their repertoire, a third class of bees included those that performed all behaviours in their repertoire at similar frequencies. There was no difference in the ages of the bees in these three behavioural classes. These results suggest that there is no evidence of task partitioning among bees performing the hygienic behaviour. The segregation observed could, however, be based on their response thresholds to the stimulus and/or on their ability to discriminate the various cues emanating from the dead brood.     

Chemical communication in Ropalidia marginata: Dufour's gland contains queen signal that is perceived across colonies and does not contain colony signal

Queens of the primitively eusocial wasp Ropalidia marginata appear to maintain reproductive monopoly through pheromone rather than through physical aggression. Upon queen removal, one of the workers (potential queen, PQ) becomes extremely aggressive but drops her aggression immediately upon returning the queen. If the queen is not returned, the PQ gradually drops her aggression and becomes the next queen of the colony. In a previous study, the Dufour's gland was found to be at least one source of the queen pheromone. Queen-worker classification could be done with 100% accuracy in a discriminant analysis, using the compositions of their respective Dufour's glands. In a bioassay, the PQ dropped her aggression in response to the queen's Dufour's gland macerate, suggesting that the queen's Dufour's gland contents mimicked the queen herself. In the present study, we found that the PQ also dropped her aggression in response to the macerate of a foreign queen's Dufour's gland. This suggests that the queen signal is perceived across colonies. This also suggests that the Dufour's gland in R. marginata does not contain information about nestmateship, because queens are attacked when introduced into foreign colonies, and hence PQ is not expected to reduce her aggression in response to a foreign queen's signal. The latter conclusion is especially significant because the Dufour's gland chemicals are adequate to classify individuals correctly not only on the basis of fertility status (queen versus worker) but also according to their colony membership, using discriminant analysis. This leads to the additional conclusion (and precaution) that the ability to statistically discriminate organisms using their chemical profiles does not necessarily imply that the organisms themselves can make such discrimination.     

Venom gland of the digger wasp Liris niger: morphology, ultrastructure, age-related changes and biochemical aspects

Females of the parasitoid digger wasp species Liris niger hunt crickets as food for their future brood. The wasps paralyse the prey by injecting their venom directly into the CNS. The venom is produced in a gland consisting of two ramified glandular tubules terminating in a common reservoir. The reservoir contents enter the sting bulb via a ductus venatus. Secretory units of dermal gland type III line the two free gland tubules, the afferent ducts to the reservoir and the cap region within the reservoir. Secretion products of tubules reach the reservoir through the cuticle-lined central funnel. Secretory cells in the distal and middle parts of the tubules contain extensive rough endoplasmic reticulum and numerous electron-dense vesicles, whereas secretory cells of the afferent ducts and the cap region of the reservoir lack electron-dense vesicles and the endoplasmic reticulum is poorly developed. The secretory apparatus undergoes age-related changes. The secretory units in the venom gland tubules and inside the reservoir complete differentiation 1 day after imaginal ecdysis. After 30 days, massive autolytic processes occur in the secretory cells and in the epithelial cells of the reservoir. Analysis of the polypeptide composition demonstrates that the venom reservoir contains numerous proteins ranging from 3.4 to 200 kDa. A dominant component is a glycoprotein of about 90 kDa. In contrast the polypeptide composition of Dufour's gland is completely different and contains no glycoproteins. Comparison of the venom reservoir contents with the polypeptide pattern of venom droplets reveals that all of the major proteinaceous constituents become secreted. Thus the secreted venom contains exclusively proteins present in the soluble contents of the venom gland.