Morphology and distribution of antennal sensilla in five species of solitary bees (Hymenoptera, Apoidea).

Solitary bees play a crucial role in ecological systems, contributing to the pollination of crops and wild plants. All females are reproductive, and their habitat requirements include nesting sites, food resources and nesting materials. Although these activities require the ability to detect biotic and abiotic stimuli in the environment, the sensory system of these species is poorly studied. In this study, the antennal sensilla of five solitary bee species belonging to three Apoidea families were investigated using scanning electron microscopy. These included two species of stem-nesting bees, Ceratina cucurbitina (Rossi, 1792) (Apidae) and Osmia scutellaris (Morawitz, 1868) (Megachilidae), and three species of ground-nesting bees, Lasioglossum brevicorne (Schenck, 1870), Lasioglossum leucozonium (Schrank, 1781), and Lasioglossum villosulum (Kirby, 1802) (Halictidae). Thirteen different types of antennal sensilla were identified in females based on their morphological characteristics: sensilla trichodea (subtypes STI, II, III), chaetica (subtypes SchI, II), basiconica (subtypes SBI, II, III, IV), placodea, campaniformia, coeloconica, and ampullacea. Their functional role was discussed and morphology was compared among the species and within the antennal segments in each species. The results provide a baseline for further physiological and behavioural studies to determine the role of antennal sensilla in habitat selection, food search and nesting site selection.

Seasonal changes in ultrastructure and gene expression in the fat body of worker honey bees.

The anatomical, physiological, and behavioral characteristics of honey bees are affected by the season as well as division of labor. In this study, we examined the structure, ultrastructure, and gene expression of fat body cells in both long-lived winter and short-lived summer worker bees (the youngest stage of hive bees and forager bees). In contrast to hive bees, foragers and winter bees have a higher metabolism due to intensive muscle activity during their flight (foragers) or endothermic heat production (winter bees). These workers differ from hive bees in the biology of their mitochondria, peroxisomes, and lysosomes as well as in the expression of the genes involved in lipid, carbohydrate, amino acid metabolism, insulin, and TGF- ß signaling. Additionally, the expression of genes related to phospholipid metabolism was higher in the hive bees. However, we found no differences between workers in the expression of genes controlling cell organelles, such as the Golgi apparatus, endoplasmic reticulum, ribosomes, nucleus, and vacuoles, as well as genes for DNA replication, cell cycle control, and autophagy. Furthermore, lysosomes, autophagic processes and lipofuscin particles were more frequently observed in winter bees using electron microscopy.

A projectome of the bumblebee central complex.

Insects have evolved diverse and remarkable strategies for navigating in various ecologies all over the world. Regardless of species, insects share the presence of a group of morphologically conserved neuropils known collectively as the central complex (CX). The CX is a navigational center, involved in sensory integration and coordinated motor activity. Despite the fact that our understanding of navigational behavior comes predominantly from ants and bees, most of what we know about the underlying neural circuitry of such behavior comes from work in fruit flies. Here, we aim to close this gap, by providing the first comprehensive map of all major columnar neurons and their projection patterns in the CX of a bee. We find numerous components of the circuit that appear to be highly conserved between the fly and the bee, but also highlight several key differences which are likely to have important functional ramifications.

Bumblebees forage widely for pollen and nectar from flowers, sometimes travelling kilometers away from their nest, but they can somehow always find their way home in a nearly straight line. These insects have been known to return to their nest from new locations almost 10 kilometers away. This homing ability is a complex neurological feat and requires the brain to combine several processes, including observing the external world, controlling bodily movements and drawing on memory. While the navigational behavior of bees has been well-studied, the neuronal circuitry behind it has not. Unfortunately, most of what is known about insects' brain activity comes from studies in species such as locusts or fruit flies. In these species, a region of the brain known as the central complex has been shown to have an essential role in homing behaviors. However, it is unknown how similar the central complex of bumblebees might be to fruit flies' or locusts', or how these differences may affect navigational abilities. Sayre et al. obtained images of thin slices of the bumblebee central complex using a technique called block-face electron microscopy, which produces high-resolution image volumes. These images were used to obtain a three-dimensional map of over 1300 neurons. This cellular atlas showed that key aspects of the central complex are nearly identical between flies and bumblebees, including the internal compass that monitors what direction the insect is travelling in. However, hundreds of millions of years of independent evolution have resulted in some differences. These were found in neurons possibly involved in forming memories of the directions and lengths of travelled paths, and in the circuits that use such vector memories to steer the insects towards their targets. Sayre et al. propose that these changes underlie bees' impressive ability to navigate. These results help explain how the structure of insects' brains can determine homing abilities. The insights gained could be used to develop efficient autonomous navigation systems, which are challenging to build and require a lot more processing power than offered by a small part of an insect brain.

Distal leg structures of the Aculeata (Hymenoptera): A comparative evolutionary study of Sceliphron (Sphecidae) and Formica (Formicidae).

The distal parts of the legs of Sceliphron caementarium (Sphecidae) and Formica rufa (Formicidae) are documented and discussed with respect to phylogenetic and functional aspects. The prolegs of Hymenoptera offer an array of evolutionary novelties, mainly linked with two functional syndromes, walking efficiently on different substrates and cleaning the body surface. The protibial-probasitarsomeral cleaning device is almost always well-developed. A complex evolutionary innovation is a triple set of tarsal and pretarsal attachment devices, including tarsal plantulae, probasitarsomeral spatulate setae, and an arolium with an internal spring-like arcus, a dorsal manubrium, and a ventral planta. The probasitarsal adhesive sole and a complex arolium are almost always preserved, whereas the plantulae are often missing. Sceliphron has retained most hymenopteran ground plan features of the legs, and also Formica, even though the adhesive apparatus of Formicidae shows some modifications, likely linked to ground-oriented habits of most ants. Plantulae are always absent in extant ants, and the arolium is often reduced in size, and sometimes vestigial. The arolium contains resilin in both examined species. Additionally, resilin enriched regions are also present in the antenna cleaners of both species, although they differ in which of the involved structures is more flexible, the calcar in Sceliphron and the basitarsal comb in Formica. Functionally, the hymenopteran distal leg combines (a) interlocking mechanisms (claws, spine-like setae) and (b) adhesion mechanisms (plantulae, arolium). On rough substrate, claws and spine-like setae interlock with asperities and secure a firm grip, whereas the unfolding arolium generates adhesive contact on smooth surfaces. Differences of the folded arolium of Sceliphron and Formica probably correlate with differences in the mechanism of folding/unfolding.

Antennal morphology and sensillar equipment vary with pollen diet specialization in Andrena bees.

Several studies recently reported that specialized (oligolectic) bees, which collect pollen from few host plants, use, besides visual cues, specific volatiles to find their hosts. Generalist (polylectic) bees, on the other hand, likely have to recognize a wider range of volatiles because they forage on many plant species. Bee antennal sensory equipment may thus be under selection to optimize plant host recognition. This selection may have led to variation in sensory equipment morphology with diet specialization (lecty). We tested if lecty correlates with antennal morphology and abundance of the main olfactory/gustatory sensilla (sensilla trichoidea (ST), placoidea (SP), sensilla basiconica (SB)) in the genus Andrena (Hymenoptera: Andrenidae). Across 24 species, and after having controlled for body size, we found polylectic species to have a longer and narrower flagellomer F9 (the one with highest abundance of sensilla), and to have a greater ST density on F9, compared with oligolectic species. Neither SP density nor SB number varied with lecty. A cluster analysis furthermore depicted groups of species that reasonably reflect diet specialization. Our results are in line with the previously observed lower number of glomeruli in the brain of oligolectic, compared with polylectic, bees. A formal correction for phylogeny is necessary to confirm our preliminary conclusion that pollen diet specialization has driven the morphology of the peripheral sensory system in this bee genus.

Structure, properties and functions of the forewing-hindwing coupling of honeybees.

Worker honeybees (Apis mellifera) are morphologically four-winged, but are functionally dipterous insects. During flight, their fore- and hindwings are coupled by means of the forewing posterior rolled margin (PRM) and hindwing hamuli. Morphological analysis shows that the PRM can be connected to the hamuli, so that the fore- and hindwing are firmly hinged, and can rotate with respect to each other. In the present study, using a combination of scanning electron microscopy (SEM) and confocal laser scanning microscopy (CLSM), we investigate the micromorphology and material composition of the coupling structures on both fore- and hindwings. High-speed filming is utilized to determine the angle variation between the fore- and hindwings in tethered flight. Using sets of two-dimensional (2D) computation fluid dynamic analyses, we further aim to understand the influence of the angle variation on the aerodynamic performance of the coupled wings. The results of the morphological investigations show that both PRM and hamuli are made up of a strongly sclerotized cuticle. The sclerotized hinge-like connection of the coupling structure allows a large angle variation between the wings (135°-235°), so that a change is made from an obtuse angle during the pronation and downstroke to a reflex angle during the supination and upstroke. Our computational results show that in comparison to a model with a rigid coupling hinge, the angle variation of a model having a flexible hinge results in both increased lift and drag with a higher rate of drag increase. This study deepens our understanding of the wing-coupling mechanism and functioning of coupled insect wings.

Honeybee pupal length assessed by CT-scan technique: effects of Varroa infestation, developmental stage and spatial position within the brood comb.

Honeybee pupae morphology can be affected by a number of stressor, but in vivo investigation is difficult. A computed tomography (CT) technique was applied to visualize a comb's inner structure without damaging the brood. The CT scan was performed on a brood comb containing pupae developed from eggs laid by the queen during a time window of 48 hours. From the CT images, the position of each pupa was determined by recording coordinates to a common reference point. Afterwards, every brood cell was inspected in order to assess the developmental stage of the pupa, the presence of Varroa destructor, the number and progeny of foundress mites. Using data on 651 pupae, the relationships between varroa infestation status, developmental stage and spatial position of the pupa within the brood comb, and its length were investigated. Pupae at 8 post-capping days were shorter than pupae at 7 post-capping days. Pupae in infected cells were significantly shorter than those in varroa-free cells and this effect was linked both to mite number and stage and to the position in the comb. Overall, the results suggest that the CT-scan may represent a suitable non-invasive tool to investigate the morphology and developing status of honeybee brood.

Kinematics of Stewart Platform Explains Three-Dimensional Movement of Honeybee's Abdominal Structure.

The Stewart platform is a typical parallel mechanism, used extensively in flight simulators with six degrees of freedom. It is rarely found in animals and has never been reported to regulate and control physiological activities. Now an equivalent Stewart platform structure is found in the honey bee (Hymenoptera: Apidae: Apis mellifera L.) abdomen to explain its three-dimensional movements. The stereoscope and scanning electron microscope are used to observe the internal structures of honeybees' abdomens. Experimental observations show that the muscles and intersegmental membranes connect the terga with the sterna and guarantee the honey bee abdominal movements. From the perspective of mechanics, a Stewart platform is evolved from the lateral connection structure of the honey bee abdomen, and the intrasegmental muscles between the sternum and tergum function as actuators between planes of the Stewart platform. The extraordinary structure provides various advantages for a honey bee to complete a variety of physiological activities. This equivalent Stewart platform structure can also be used to illustrate the flexible abdominal movements of other insects with the segmental abdomen.

A comparative study of the pharyngeal plate of Apoidea (Hymenoptera: Aculeata), with implications for the understanding of phylogenetic relationships of bees.

The pharyngeal plate is a morphological complex with extensive anatomical variation among bees and, therefore, potential as a source of phylogenetic information. The pharyngeal plate of bees is divided into four morphologically distinct regions: sitophore, hypopharyngeal lobe, pharyngeal rods, and median oral plate. In this work we illustrate and document in detail for the first time the pharyngeal plate of 43 bee species, providing descriptions of the morphological variation and contrasting these findings with representatives of apoid wasps (Crabronidae and Sphecidae). We evaluate and discuss the potential of this structure as a rich source of morphological information in the context of bee phylogeny and any research potentially impacted by comparative morphological data. The shape of the hypopharyngeal lobe is highly variable among suprageneric taxa of bees and can be readily employed to characterise taxa at various levels. We argue that the global patterns in the variation of the pharyngeal plate can provide information for phylogenetic inference within bees and constructed and coded 10 characters that encompass the most noticeable morphological differences discussed herein.

Using micro-CT techniques to explore the role of sex and hair in the functional morphology of bumblebee (Bombus terrestris) ocelli.

Many insects have triplets of camera type eyes, called ocelli, whose function remains unclear for most species. Here, we investigate the ocelli of the bumblebee, Bombus terrestris, using reconstructed 3D data from X-ray microtomography scans combined with computational ray-tracing simulations. This method enables us, not only to predict the visual fields of the ocelli, but to explore for the first time the effect that hair has on them as well as the difference between worker female and male ocelli. We find that bumblebee ocellar fields of view are directed forward and dorsally, incorporating the horizon as well as the sky. There is substantial binocular overlap between the median and lateral ocelli, but no overlap between the two lateral ocelli. Hairs in both workers and males occlude the ocellar field of view, mostly laterally in the worker median ocellus and dorsally in the lateral ocelli. There is little to no sexual dimorphism in the ocellar visual field, suggesting that in B. terrestris they confer no advantage to mating strategies. We compare our results with published observations for the visual fields of compound eyes in the same species as well as with the ocellar vision of other bee and insect species.

Physical immobilization of particles inspired by pollination.

Biomimetic systems often exhibit striking designs well adapted to specific functions that have been inspiring the development of new technologies. Herein, we explored the remarkable ability of honey bees to catch and release large quantities of pollen grains. Hair spacing and height on bees are crucial for their ability to mechanically fix pollen grains. Inspired by this, we proposed the concept of a micropatterned surface for microparticle entrapment, featuring high-aspect-ratio elastic micropillars spaced to mimic the hairy surface of bees. The hypothesis was validated by investigating the ability of polydimethylsiloxane microfabricated patches to fix microparticles. The geometrical arrangement, spacing, height, and flexibility of the fabricated micropillars, and the diameter of the microparticles, were investigated. Higher entrapment capability was found through the match between particle size and pillar spacing, being consistent with the observations that the diameter of pollen grains is similar to the spacing between hairs on bees' legs. Taller pillars permitted immobilization of higher quantities of particles, consistent with the high aspect ratio of bees' hairs. Our biomimetic surfaces were explored for their ability to fix solid microparticles for drug-release applications, using tetracycline hydrochloride as a model antibiotic. These surfaces allowed fixation of more than 20 mg/cm2 of antibiotic, about five times higher dose than commercialized patches (5.1 mg/cm2). Such bioinspired hairy surfaces could find applications in a variety of fields where dry fixation of high quantities of micrometer-sized objects are needed, including biomedicine, agriculture, biotechnology/chemical industry, and cleaning utensils.

Phylogenetic analyses of antibiotic-producing Streptomyces sp. isolates obtained from the stingless-bee Tetragonisca angustula (Apidae: Meliponini).

Many insects have been associated with actinobacteria in protective symbiosis where antimicrobial metabolites inhibit host pathogens. However, the microbiota of neotropical insects such as the stingless-bee Tetragonisca angustula is poorly explored. T. angustula is a meliponid bee widely distributed in Latin America, its honey is traditionally exploited because of its ethno-pharmacological properties and its antimicrobial activity has been demonstrated. Also, the well-structured nest of this species allows exploration of the microbiota of its different components. Even though Streptomyces spp. have been cultured from stingless-bees, little is known about their role in this insect-microbe relationship. In this study, we examined the association between culturable actinobacteria and T. angustula, and evaluated the isolates' potential as antimicrobial producers. We isolated 51 actinobacteria from adult bees and different substrates of the hive of T. angustula (pollen and honey storage, garbage pellets and cerumen). We then performed a 16S rRNA phylogenetic analysis that clusters the bacteria to previously described lineages of host-associated Streptomyces. In addition, all the isolates were classified according to their antibacterial activity against human pathogens, measured by a growth inhibition test based on diffusion in agar. More than 50 % of our isolates exhibit antimicrobial activity, mainly to Gram-positive bacteria and fungi and only two against Gram-negative bacteria. Additionally, we obtained electron micrographs of adult bees with what appears to be patches of hyphae with Streptomyces-like cell morphology on their body surface. Our results suggest that T. angustula possibly uptakes and transfers actinobacteria from the environment, acting as vectors for these potentially beneficial organisms. This research provides new insights regarding the microbiota associated with T. angustula and justify future studies exploring the full diversity of the microbial community associated with the hive and the possible exchange of microbes with the crops they pollinate.

Post-embryonic development of the Malpighian tubules in Apis mellifera (Hymenoptera) workers: morphology, remodeling, apoptosis, and cell proliferation.

The honeybee Apis mellifera has ecological and economic importance; however, it experiences a population decline, perhaps due to exposure to toxic compounds, which are excreted by Malpighian tubules. During metamorphosis of A. mellifera, the Malpighian tubules degenerate and are formed de novo. The objective of this work was to verify the cellular events of the Malpighian tubule renewal in the metamorphosis, which are the gradual steps of cell remodeling, determining different cell types and their roles in the excretory activity in A. mellifera. Immunofluorescence and ultrastructural analyses showed that the cells of the larval Malpighian tubules degenerate by apoptosis and autophagy, and the new Malpighian tubules are formed by cell proliferation. The ultrastructure of the cells in the Malpighian tubules suggest that cellular remodeling only occurs from dark-brown-eyed pupae, indicating the onset of excretion activity in pupal Malpighian tubules. In adult forager workers, two cell types occur in the Malpighian tubules, one with ultrastructural features (abundance of mitochondria, vacuoles, microvilli, and narrow basal labyrinth) for primary urine production and another cell type with dilated basal labyrinth, long microvilli, and absence of spherocrystals, which suggest a role in primary urine re-absorpotion. This study suggests that during the metamorphosis, Malpighian tubules are non-functional until the light-brown-eyed pupae, indicating that A. mellifera may be more vulnerable to toxic compounds at early pupal stages. In addition, cell ultrastructure suggests that the Malpighian tubules may be functional from dark-brown-eyed pupae and acquire greater complexity in the forager worker bee.

Drinking with a very long proboscis: Functional morphology of orchid bee mouthparts (Euglossini, Apidae, Hymenoptera).

Neotropical orchid bees (Euglossini) possess the longest proboscides among bees. In this study, we compared the feeding behavior and functional morphology of mouthparts in two similarly large-sized species of Euglossa that differ greatly in proboscis length. Feeding observations and experiments conducted under semi-natural conditions were combined with micro-morphological examination using LM, SEM and micro CT techniques. The morphometric comparison showed that only the components of the mouthparts that form the food tube differ in length, while the proximal components, which are responsible for proboscis movements, are similar in size. This study represents the first documentation of lapping behaviour in Euglossini. We demonstrate that Euglossa bees use a lapping-sucking mode of feeding to take up small amounts of fluid, and a purely suctorial technique for larger fluid quantities. The mouthpart movements are largely similar to that in other long-tongued bees, except that the postmentum in Euglossa can be extended, greatly enhancing the protraction of the glossa. This results in a maximal functional length that is about 50% longer than the length of the food canal composing parts of the proboscis. The nectar uptake and the sensory equipment of the proboscis are discussed in context to flower probing.

Survey of Hatching Spines of Bee Larvae Including Those of Apis mellifera (Hymenoptera: Apoidea).

This article explores the occurrence of hatching spines among bee taxa and how these structures enable a larva on hatching to extricate itself from the egg chorion. These spines, arranged in a linear sequence along the sides of the first instar just dorsal to the spiracles, have been observed and recorded in certain groups of solitary and cleptoparasitic bee taxa. After eclosion, the first instar remains loosely covered by the egg chorion. The fact that this form of eclosion has been detected in five families (Table 1 identifies four of the families. The fifth family is the Andrenidae for which the presence of hatching spines in the Oxaeinae will soon be announced.) of bees invites speculation as to whether it is a fundamental characteristic of bees, or at least of solitary and some cleptoparasitic bees. The wide occurrence of these spines has prompted the authors to explore and discover their presence in the highly eusocial Apis mellifera L. Hatching spines were indeed discovered on first instar A. mellifera. The honey bee hatching process appears to differ in that the spines are displayed somewhat differently though still along the sides of the body, and the chorion, instead of splitting along the sides of the elongate egg, seems to quickly disintegrate from the emerging first instar in association with the nearly simultaneous removal of the serosa that covers and separates the first instar from the chorion. Unexpected observations of spherical bodies of various sizes perhaps containing dissolving enzymes being discharged from spiracular openings during hatching may shed future light on the process of how A. mellifera effects chorion removal during eclosion. Whereas hatching spines occur among many groups of bees, they appear to be entirely absent in the Nomadinae and parasitic Apinae, an indication of a different eclosion process.

Post-embryonic changes in the hindgut of honeybee Apis mellifera workers: Morphology, cuticle deposition, apoptosis, and cell proliferation.

In insects, the hindgut is a homeostatic region of the digestive tract, divided into pylorus, ileum, and rectum, that reabsorbs water, ions, and small molecules produced during hemolymph filtration. The hindgut anatomy in bee larvae is different from that of adult workers. This study reports the morphological changes and cellular events that occur in the hindgut during the metamorphosis of the honeybee Apis mellifera. We describe the occurrence of autophagosomes and the ultrastructure of the epithelial cells and cuticle, suggesting that cuticular degradation begins in prepupae, with the cuticle being reabsorbed and recycled by autophagosomes in white- and pink-eyed pupae, followed by the deposition of new cuticle in light-brown-eyed pupae. In L5S larvae and prepupae, the hindgut undergoes cell proliferation in the anterior and posterior ends. In the pupae, the pylorus, ileum, and rectum regions are differentiated, and cell proliferation ceases in dark-brown-eyed pupae. Apoptosis occurs in the hindgut from the L5S larval to the pink-eyed pupal stage. In light-brown- and dark-brown-eyed pupae, the ileum epithelium changes from pseudostratified to simple only after the production of the basal lamina, whereas the rectal epithelium is always flattened. In black-eyed pupae, ileum epithelial cells have large vacuoles and subcuticular spaces, while in adult forager workers these cells have long invaginations in the cell apex and many mitochondria, indicating a role in the transport of compounds. Our findings show that hindgut morphogenesis is a dynamic process, with tissue remodeling and cellular events taking place for the formation of different regions of the organ, the reconstruction of a new cuticle, and the remodeling of visceral muscles.

Three-dimensional visualization of ocellar interneurons of the orchid bee Euglossa imperialis using micro X-ray computed tomography.

We used contrast-optimized micro X-ray computed tomography (mCT) to trace the profiles of the full complement of large ocellar L-neurons in the male orchid bee Euglossa imperialis. We find that most L-neurons collect information from either the dorsal or the ventral retinae in both median and lateral ocelli, with only three neurons associated with the median ocellus having dendritic branches in both dorsal and ventral retina. In the median ocellus, we find also L-neurons that either collect information from the left or the right half of the ocellar plexus and two neurons that have a split dendritic tree in both halves. Fourteen large L-neurons collect information from the median ocellus and six to seven L-neurons from each of the lateral ocelli. The only L-neurons that project to the contralateral protocerebrum are those that have their dendritic branches in the ventral plexi of both median and lateral ocelli. The target areas of dorsal L-neurons from the lateral ocelli include a tract of mechanosensory fibers originating in the antennae. We compare our findings with what is known from the ocellar systems of other insects, make a number of functional inferences and discuss the advantages and disadvantages of mCT scans for the purpose of tracing large neuron profiles.

Development of antennal sensilla of Tetragonisca angustula Latreille, 1811 (Hymenoptera: Meliponini) during pupation / Desenvolvimento das sensilas antenais de Tetragonisca angustula latreille, 1811 (Hymenoptera: Meliponini) durante a pupação

Abstract The antennal sensilla are sensory organs formed by a group of neurons and accessory cells, which allow perception of environmental cues, which play a role as mechanoreceptors and chemoreceptors. This study describes the post-embryonic development of the antennal sensilla of the stingless Tetragonisca angustula (Hymenoptera: Meliponini) workers. The development of the antennal sensilla begins in the transition stage of the pre-pupae to white-eyed pupae. The sensilla are completely developed at the black-eyed pupae stage, but they are covered by the old cuticle. The sensilla are exposed to the environment only in newly emerged workers of T. angustula, but it is possible that environmental stimuli can be recognized due to the pores in the old cuticle.

Resumo As sensilas antenais são órgãos sensoriais formados por um conjunto de neurônios que captam estímulos ambientais e células acessórias, desempenhando as funções de mecanorreceptores e quimiorrecepterores. Este trabalho descreve o desenvolvimento pós-embrionário das sensilas antenais de operárias das abelhas sem ferrão Tetragonisca angustula (Hymenoptera: Meliponini). O desenvolvimento das sensilas tem início na transição de pré-pupa para pupa de olho branco e estão completamente desenvolvidas no estágio de pupa de olho preto, mas ainda estão cobertas pela cutícula velha. As sensilas estão completamente expostas em operárias recém-emergidas de T. angustula, mas é possível que estímulos ambientais sejam percebidos em estágios anteriores devido aos poros presentes na cutícula velha.

Characterization of antennal sensilla, larvae morphology and olfactory genes of Melipona scutellaris stingless bee.

There is growing evidence in the literature suggesting that caste differentiation in the stingless bee, Melipona scutellaris, and other bees in the genus Melipona, is triggered by environmental signals, particularly a primer pheromone. With the proper amount of food and a chemical stimulus, 25% of females emerge as queens, in agreement with a long-standing "two loci/two alleles model" proposed in the 1950s. We surmised that these larvae must be equipped with an olfactory system for reception of these chemical signals. Here we describe for the first time the diversity of antennal sensilla in adults and the morphology of larvae of M. scutellaris. Having found evidence for putative olfactory sensilla in larvae, we next asked whether olfactory proteins were expressed in larvae. Since the molecular basis of M. scutellaris is still unknown, we cloned olfactory genes encoding chemosensory proteins (CSP) and odorant-binding proteins (OBPs) using M. scutellaris cDNA template and primers designed on the basis CSPs and OBPs previously reported from the European honeybee, Apis mellifera. We cloned two CSP and two OBP genes and then attempted to express the proteins encoded by these genes. With a recombinant OBP, MscuOBP8, and a combinatorial single-chain variable fragment antibody library, we generated anti-MscuOBP8 monoclonal antibody. By immunohistochemistry we demonstrated that the anti-MscuOBP8 binds specifically to the MscuOBP8. Next, we found evidence that MscuOBP8 is expressed in M. scutellaris larvae and it is located in the mandibular region, thus further supporting the hypothesis of olfactory function in immature stages. Lastly, molecular modeling suggests that MscuOBP8 may function as a carrier of primer pheromones or other ligands.

X-ray micro computed-tomography.

Emily Baird and Gavin Taylor describe how you can make three-dimensional models of biological samples using x-ray micro-computed tomography.