Modifications in the oviducts of workers and queens of <Emphasis Type="Italic">Melipona quadrifasciata anthidioides</Emphasis> (Hymenoptera: Apidae) with different ages

The study of morphological features of highly eusocial bees has helped to answer a series of questions concerning their biology. The labor division places the females into two castes, queen with reproductive function and worker with a wide variety of tasks. However, in different species and under different conditions, workers can develop ovaries and lay trophic eggs that are eaten by the queen or used to originate males. In this work, the development of the oviducts was monitored in workers and virgin queens of Melipona quadrifasciata anthidioides specimens of different ages to verify whether there is permanent sterility of these queens and workers due to aging. Lateral and common oviducts of virgin queens aged <7, 10, 15, 20, and 25 days old, physogastric queens, nurse, and forager workers were analyzed for histology and ultrastructure. Although the structural organization of the lateral and common oviducts were very similar, differences in width of the cuticle and the degree of chromatin condensation were observed, indicating differences in the development of this organ of the reproductive system between the castes. It was also demonstrated that electron-lucent vesicles appear to be related to the formation of the cuticle that lines the oviducts. Because no cellular death characteristics were found, it can be inferred that the absence of mating of the queens as old as of 25 days of age does not cause permanent sterility.     

Occurrence and morphometry of the hypopharyngeal glands inScaptotrigona postica Lat. (Hymenoptera, Apidae, Meliponinae)

As compared toApis mellifera where only workers have hypopharyngeal glands, inScaptotrigona postica, these glands occur in workers, queens and males. They are composed of two long axial ducts with many unicellular secretory alveoli interconnected by secretory canaliculi. The axial ducts are longer in males than in workers, but the alveolar areas of queens and males are generally smaller. In workers the alveoli have their greatest size in the nurses or middle-aged individuals while in queens and males they are larger in newly emerged individuals. The results indicate that the glands in workers may produce food for the brood as inA. mellifera, since they are well developed in the nurse workers. However, the function of the glands in queens and males remains to be clarified since these individuals have no part in brood care.     

Nestmate Recognition Differences between Honeybee Colonies of <Emphasis Type="BoldItalic">Apis cerana</Emphasis> and <Emphasis Type="BoldItalic">Apis mellifera</Emphasis>

Nestmate recognition in Apis cerana and Apis mellifera was studied by introducing sealed queen cells heterospecifically between queenless colonies. No A. cerana queens were accepted by queenless A. mellifera; but A. mellifera queens were accepted in queenless A. cerana colonies. A. mellifera queens oviposited in queenless A. cerana colonies, but A. cerana workers removed most eggs. In time, egg removals declined, and some A. mellifera larvae that hatched from these eggs reached adulthood, and eventually about half of the workers were newly emerged A. mellifera. Eventually, the colonies consisted only of A. mellifera after A. cerana workers died by attrition. A. mellifera workers are more sensitive to nestmate recognition and killed the A. cerana virgin queens. In mixed-species colonies, after newly emerged A. mellifera workers matured, they removed eggs laid by the A. cerana queens until there were no workers to replace the old ones.     

Caste- and sex-specific adaptations within the olfactory pathway in the brain of the ant Camponotus floridanus

Olfaction plays a key role in mediating ant behavior, and ant societies are characterized by caste- and sex-specific division of labor. We propose that caste- and sex-specific adaptations in the olfactory pathway promote differences in olfactory behavior. This study compares olfactory centers in the brain of large (major) workers, small (minor) workers, virgin queens, and males of the carpenter ant Camponotus floridanus. The number of glomeruli in the antennal lobe was similar in the female castes, although the glomerular volumes differed. Males had approximately 45% fewer glomeruli compared to females (approximately 258 and approximately 434) and one antennal sensory tract was absent. A dual output pathway to the mushroom bodies was present in males. In contrast to females, however, the number of glomeruli connected to the medial antennocerebral tract was substantially smaller than those associated with the lateral tract. All glomeruli in the male antennal lobe contained serotonergic processes, whereas in the female castes glomeruli in the large tract six cluster lacked serotonergic innervations. We conclude that differences in general glomerular organization are subtle among the female castes, but sex-specific differences in the number, connectivity and neuromodulatory innervation of glomeruli are substantial and likely to underlie differences in olfactory processing and learning.     

Host choice in the phoretic mite Parasitellus fucorum (Mesostigmata: Parasitidae): which bumblebee caste is the best?

Host caste recognition may be important for the dispersal of phoretic mites associated with social insects. All developmental stages of the mite Parasitellus fucorum (Acari: Mesostigmata: Parasitidae) live in the nests of bumblebees (Hymenoptera: Apidae: Bombus). Dispersal occurs by specialised phoretic instars, deutonymphs, which attach to adult bumblebees. Since bumblebee colonies are annual and only young queens overwinter, deutonymphs that are able to discriminate between bumblebee castes and preferentially attach to queens should be favoured by selection. In the field, deutonymphs of P. fucorum were found to be phoretic on bumblebee workers and queens, and in behavioural experiments all castes proved to be attractive as carriers for the mites. However, they preferred queens that had hibernated as carriers when they could choose between workers and queens. In a further experiment, when given a choice, deutonymphs switched from males to young queens but never transferred from a queen to a male. These results suggest that deutonymphs preferentially attach to queens but may also use other castes for transport. Those dispersing on workers and males may try to switch to queens later. Host-switching is possible during copulation and on flowers, where bees of all castes forage. Received: 14 November 1997 / Accepted: 16 February 1998     

Ergatoid queen development in the ant Myrmecina nipponica: modular and heterochronic regulation of caste differentiation

Caste polyphenism in social insects provides us with excellent opportunities to examine the plasticity and robustness underlying developmental pathways. Several ant species have evolved unusual castes showing intermediate morphologies between alate queens and wingless workers. In some low-temperature habitats, the ant Myrmecina nipponica produces such intermediate reproductives (i.e. ergatoids), which can mate and store sperm but cannot fly. To gain insight into the developmental and evolutionary aspects associated with ergatoid production, we conducted morphological and histological examinations of the post-embryonic development of compound eyes, gonads and wings during the process of caste differentiation. In compound eyes, both the queen-worker and ergatoid-worker differences were already recognized at the third larval instar. In gonads, queen-worker differentiation began at the larval stage, and ergatoid-worker differentiation began between the prepupal and pupal stages. Wing development in ergatoids was generally similar to that in workers throughout post-embryonic development. Our results showed that the developmental rate and timing of differentiation in body parts differed among castes and among body parts. These differences suggest that the rearrangement of modular body parts by heterochronic developmental regulation is responsible for the origination of novel castes, which are considered to be adaptations to specific ecological niches.     

Gene expression patterns associated with caste and reproductive status in ants: worker‐specific genes are more derived than queen‐specific ones

Variation in gene expression leads to phenotypic diversity and plays a central role in caste differentiation of eusocial insect species. In social Hymenoptera, females with the same genetic background can develop into queens or workers, which are characterized by divergent morphologies, behaviours and lifespan. Moreover, many social insects exhibit behaviourally distinct worker castes, such as brood‐tenders and foragers. Researchers have just started to explore which genes are differentially expressed to achieve this remarkable phenotypic plasticity. Although the queen is normally the only reproductive individual in the nest, following her removal, young brood‐tending workers often develop ovaries and start to reproduce. Here, we make use of this ability in the ant Temnothorax longispinosus and compare gene expression patterns in the queens and three worker castes along a reproductive gradient. We found the largest expression differences between the queen and the worker castes (~2500 genes) and the smallest differences between infertile brood‐tenders and foragers (~300 genes). The expression profile of fertile workers is more worker‐like, but to a certain extent intermediate between the queen and the infertile worker castes. In contrast to the queen, a high number of differentially expressed genes in the worker castes are of unknown function, pointing to the derived status of hymenopteran workers within insects.     

Making a queen: an epigenetic analysis of the robustness of the honeybee (Apis mellifera) queen developmental pathway

Specialized castes are considered a key reason for the evolutionary and ecological success of the social insect lifestyle. The most essential caste distinction is between the fertile queen and the sterile workers. Honeybee (Apis mellifera) workers and queens are not genetically distinct, rather these different phenotypes are the result of epigenetically regulated divergent developmental pathways. This is an important phenomenon in understanding the evolution of social insect societies. Here, we studied the genomic regulation of the worker and queen developmental pathways, and the robustness of the pathways by transplanting eggs or young larvae to queen cells. Queens could be successfully reared from worker larvae transplanted up to 3 days age, but queens reared from older worker larvae had decreased queen body size and weight compared with queens from transplanted eggs. Gene expression analysis showed that queens raised from worker larvae differed from queens raised from eggs in the expression of genes involved in the immune system, caste differentiation, body development and longevity. DNA methylation levels were also higher in 3‐day‐old queen larvae raised from worker larvae compared with that raised from transplanted eggs identifying a possible mechanism stabilizing the two developmental paths. We propose that environmental (nutrition and space) changes induced by the commercial rearing practice result in a suboptimal queen phenotype via epigenetic processes, which may potentially contribute to the evolution of queen–worker dimorphism. This also has potentially contributed to the global increase in honeybee colony failure rates.     

Morphological changes in the cephalic salivary glands of females and males of <Emphasis Type="Italic">Apis mellifera</Emphasis> and <Emphasis Type="Italic">Scaptotrigona postica</Emphasis> (Hymenoptera,Apidae)

The cephalic salivary glands of some species of bees are exclusive and well developed only in Apinae. These glands were studied with light and scanning electron microscopy in workers, queens and males from the honey bee Apis mellifera, and the stingless bee Scaptotrigona postica in different life phases. The results show that the cephalic salivary glands are present in females of both the species, and in males of S. postica. Nevertheless, they are poorly developed in young males of A. mellifera. In both species, gland growth is progressive from the time of emergence to the oldest age but, in A. mellifera males, the gland degenerates with age. Scanning electron microscopy shows that the secretory units of newly emerged workers are collapsed while in older workers they are turgid. Some pits on the surface of the secretory units correspond to open intercellular spaces. The possible functions of these glands in females and males of both species are discussed.     

Reproductive and worker castes in the primitively eusocial wasp Belonogaster petiolata (DeGeer) (Hymenoptera: Vespidae): evidence for pre-imaginal differentiation

Until recently, morphological differences between castes in independent-founding polistine wasps were considered absent. This paper investigates the extent of morphological and physiological differences between reproductive (foundress and gyne) and worker castes of Belonogaster petiolata, and tests the hypothesis that caste differentiation in this species occurs pre-imaginally.Foundresses were significantly larger than workers, to the extent that foundress/worker ratios were comparable with those between queens and workers in some swarm-founding Polistinae. Early emerging workers were small, but body size increased over the colony cycle such that late-season workers were similar in size to gynes. In proportion to body size, workers possessed broader heads while foundresses and gynes had broader thoraces and gasters. All queens, 98% of subordinate foundresses, and 95% of over-wintering gynes were inseminated. Workers were never inseminated and lacked mature ovaries in colonies with active queens. However, in the absence of the queen (and other foundresses), 11% of workers developed mature ovaries. Ovarian size and fat content of foundresses and gynes was significantly greater than that of workers. The differences in external morphology and reproductive physiology between castes support the hypothesis that differentiation occurs pre-imaginally. However, imaginal factors, in particular social dominance of the queen, maintain the reproductive subordinance of workers.     

Caste-Specific Differences in Hindgut Microbial Communities of Honey Bees (Apis mellifera)

Host-symbiont dynamics are known to influence host phenotype, but their role in social behavior has yet to be investigated. Variation in life history across honey bee (Apis mellifera) castes may influence community composition of gut symbionts, which may in turn influence caste phenotypes. We investigated the relationship between host-symbiont dynamics and social behavior by characterizing the hindgut microbiome among distinct honey bee castes: queens, males and two types of workers, nurses and foragers. Despite a shared hive environment and mouth-to-mouth food transfer among nestmates, we detected separation among gut microbiomes of queens, workers, and males. Gut microbiomes of nurses and foragers were similar to previously characterized honey bee worker microbiomes and to each other, despite differences in diet, activity, and exposure to the external environment. Queen microbiomes were enriched for bacteria that may enhance metabolic conversion of energy from food to egg production. We propose that the two types of workers, which have the highest diversity of operational taxonomic units (OTUs) of bacteria, are central to the maintenance of the colony microbiome. Foragers may introduce new strains of bacteria to the colony from the environment and transfer them to nurses, who filter and distribute them to the rest of the colony. Our results support the idea that host-symbiont dynamics influence microbiome composition and, reciprocally, host social behavior.     

Unique habits of stomodeal trophallaxis in the ponerine antHypoponera sp.

Summary The present paper describes for the first time that adult-to-adult trophallaxis exists in ponerine ants. Furthermore, it shows that the adult castes of the ponerine andHypoponera sp. have different habits of trophallaxis. Workers display a soliciting behavior toward queens, males and workers, but receive regurgitated food only from workers. The workers are forced to stop soliciting for regurgitated food by the whipping behavior of the queen. Callow queens solicit regurgitated food from workers and ergatoid males and receive it, while mature queens do not solicit regurgitation from workers. Ergatoid males receive regurgitated food from workers and regurgitate it to queens. Alate males show no trophallactic food exchange with workers and queens. Trophallactic behavior was never observed between males of either winged or ergatoid types.     

Morphological caste and sex differences in the Taiwanese stingless bee Trigona ventralis hoozana (Hymenoptera: Apidae)

To examine morphological differences among queens, workers and males, 14 external body characters were measured in two colonies of the Taiwanese stingless bee Trigona ventralis hoozana. Queens were largest in all of the body parts measured except eye width and mesoscutum length, and values for most variables in queens did not overlap with those of workers and males. In contrast, the worker : male size ratios for 11 variables were close to 1.0, showing that overall body size and shape of workers resembled that of males rather than of queens. Males had the largest eyes and their mesoscutum length was comparable to that of queens. ancova between 14 morphometric variables and mesoscutum width chosen as standard body size showed that allometric growth in most variables was not linear. Plotting of some variables on mesoscutum width showed that queens had a proportionally wider first metasomal tergum and longer antennal scape, but a proportionally narrower head and eye than workers and males. These tests suggest that the morphological caste differences in this species belong to a category of complete dimorphism.     

Comparative morphology of cephalic exocrine glands among castes of the black ant Lasius niger

The glandular system is crucially involved in main aspects of ant social life. The function of glands has been primarily studied in the workers (the non-reproductive individuals in a colony). In contrast, little information is available on queens (the reproductive females in a colony) or males in spite of the obvious functional differences between these castes. Here we report a comparison of the general morphology of the mandibular, propharyngeal and postpharyngeal glands between the three castes of the black ant Lasius niger. The analysis clearly shows that all these cephalic glands differ in relative size between castes and suggests a link between gland structure and its behavioral role in queens, workers and males. In particular, males present a hypertrophied mandibular gland. This is consistent with the fact that these glands might be the source of the sex pheromone in this caste. By contrast, queens exhibited the most developed postpharyngeal glands. This is consistent with the production of particular cues by queens for workers to help them to distinguish between reproductive and non-reproductive females. Finally, the propharyngeal glands were most developed in the worker caste and of similar relative size in males and queens. Their function is still enigmatic.     

Queen–worker caste ratio depends on colony size in the pharaoh ant (<Emphasis Type="Italic">Monomorium pharaonis</Emphasis>)

The success of an ant colony depends on the simultaneous presence of reproducing queens and non-reproducing workers in a ratio that will maximize colony growth and reproduction. Despite its presumably crucial role, queen–worker caste ratios (the ratio of adult queens to workers) and the factors affecting this variable remain scarcely studied. Maintaining polygynous pharaoh ant (Monomorium pharaonis) colonies in the laboratory has provided us with the opportunity to experimentally manipulate colony size, one of the key factors that can be expected to affect colony level queen–worker caste ratios and body size of eclosing workers, gynes and males. We found that smaller colonies produced more new queens relative to workers, and that these queens and workers both tended to be larger. However, colony size had no effect on the size of males or on the sex ratio of the individuals reared. Furthermore, for the first time in a social insect, we confirmed the general life history prediction by Smith and Fretwell (Am Nat 108:499–506, 1974) that offspring number varies more than offspring size. Our findings document a high level of plasticity in energy allocation toward female castes and suggest that polygynous species with budding colonies may adaptively adjust caste ratios to ensure rapid growth.     

Caste‐biases in gene expression are specific to developmental stage in the ant Formica exsecta

Understanding how a single genome creates and maintains distinct phenotypes is a central goal in evolutionary biology. Social insects are a striking example of co‐opted genetic backgrounds giving rise to dramatically different phenotypes, such as queen and worker castes. A conserved set of molecular pathways, previously envisioned as a set of ‘toolkit’ genes, has been hypothesized to underlie queen and worker phenotypes in independently evolved social insect lineages. Here, we investigated the toolkit from a developmental point of view, using RNA‐Seq to compare caste‐biased gene expression patterns across three life stages (pupae, emerging adult and old adult) and two female castes (queens and workers) in the ant Formica exsecta. We found that the number of genes with caste‐biased expression increases dramatically from pupal to old adult stages. This result suggests that phenotypic differences between queens and workers at the pupal stage may derive from a relatively low number of caste‐biased genes, compared to higher number of genes required to maintain caste differences at the adult stage. Gene expression patterns were more similar among castes within developmental stages than within castes despite the extensive phenotypic differences between queens and workers. Caste‐biased expression was highly variable among life stages at the level of single genes, but more consistent when gene functions (gene ontology terms) were investigated. Finally, we found that a large part of putative toolkit genes were caste‐biased at least in some life stages in F. exsecta, and the caste‐biases, but not their direction, were more often shared between F. exsecta and other ant species than between F. exsecta and bees. Our results indicate that gene expression should be examined across several developmental stages to fully reveal the genetic basis of polyphenisms.     

Morpho-functional comparison of the Dufour gland in the female castes of the Amazon ant <Emphasis Type="Italic">Polyergus rufescens</Emphasis> (Hymenoptera,Formicidae)

The Dufour gland is crucially involved in main aspects of the parasite habit of the slave-making ant Polyergus rufescens, i.e. slave-raids and host colony usurpation. Workers use chemicals from this gland as recruitment signals during raid organization, while newly-mated queens use its secretion to appease residents during host nest invasion. Here, we report a comparison of the general morphology and fine structure of the Dufour gland in the female castes of P. rufescens: queens, ergatogynes (intermediate forms), and workers. The analysis clearly shows the link between gland structure and its behavioural role in queens and workers. In particular, queens present a hypertrophied gland with an extended lumen and a thin epithelium no more active in secretory function. This is consistent with the fact that usurper queens use the Dufour gland contents only during the short phase of host nest penetration. Contrary to adult queens, the cytoplasmic organization of the Dufour gland epithelium of raiders is typical for a tissue with secretory activity (abundance of mitochondria, free ribosomes, strands of smooth endoplasmic reticulum and a Golgi apparatus). This is consistent with the continuous raiding activity performed by workers throughout their adult life. The biology of ergatogynes is still an enigmatic matter. Their Dufour gland is intermediate in shape and size between that of queens and workers. It presents a fairly thick epithelium with features that are typical of a quite active secretory tissue.