Electrophoretical polypeptide pattern of hypopharyngeal glands extracts from workers and males of Scaptotrigona postica Latr. (Hymenoptera, Apidae, Meliponinae)

In S. postica the hypopharyngeal glands are present in workers, males and queens. The glands of newly emerged workers are in a prefunctional phase, in nurse workers they reach their highest development and in forager workers they suffer reabsorption. The newly emerged males and queens, however, have well developed glands that soon start involution. The electrophoretical pattern of worker and male hypopharyngeal glands is compared with the pattern of nurse workers of A. mellifera. The results show simularities, as well as differences among the S. postica males and workers, and among the phases of worker life. The electrophoretic band pattern suggests that the hypopharyngeal glands of nurse workers of S. postica produce substances similar to the ones produced by A. mellifera. The similarity of the extracts of nurse glands of A. mellifera and S. postica, as well as the behavioural similarities of the workers, suggest the same function of this gland in both species.

Behavioral responses evoked in honey bee workers by Dufour gland extracts (Hymenoptera : Apidae)

A series of experiments was performed to assess possible roles of the honey bee Dufour gland secretion. Bioassays with extracts of queen and worker glands from two colonies were made under artificial conditions, in which nestmate and non-nestmate forager workers were tested. The results demonstrate that forager workers display behavioral responses when exposed to Dufour gland extracts of nestmates, but remain indifferent when exposed to non-nestmate extracts. Also, the results demonstrate that forager workers are attracted by virgin queen gland extracts, and repelled by forager worker extracts. The data demonstrate that the Dufour gland secretion is colony- and caste-specific. The attractant remains in the nest. The repellent effect of forager worker extracts is interpreted as an alarm-like pheromone. The attractant effect of virgin queen extracts could be useful in the swarming process to attract scout bees to the eventual founder virgin queen or to attract old foragers to the virgin queen that remains in the nest.

Morphological modifications induced by an artificial diet on the hypopharyngeal glands of Apis mellifera (Hymenoptera, apidae) during their degenerative process

The morphological aspects of the hypopharyngeal glands were analyzed in worker bees of Apis mellifera of 15 and 30 days of age. The individuals were kept in a room with controlled temperature at 32degreesC where they received water and either a protein or a high energy food. Nurse and foraging workers were used as a control for the experiment. The morphological results showed that the different diets modified the cell death characteristics and intensified its occurrence. Both diets caused precocious glandular degeneration. However, this anticipation of cell death was more pronounced in the glandular tissue of the workers who received the high energy diet when compared to the glands of the bees fed with the protein meal.The degenerative signs observed were an intense cytoplasmic vacuolization, with a loss of cytoplasm and of the cell boundaries, dilation or condensation of the cells and nuclei, and nuclear fragmentation. At the end of the degenerative process, we observed the extrusion of nuclei and, finally, the dissolution of the glands. The hypopharyngeal glands' remains were found in the haemolymph.

Ultrastructure of the ducts of the reproductive tract of males of Melipona bicolor bicolor Lepeletier (Hymenoptera, Apinae, Meliponini)

The present paper describes the ultrastructural features of seminal vesicle, post-vesicular vas deferens and ejaculatory duct of Melipona bicolor bicolor from newly emerged and mature males. Although the results do not show very consistent morphological signs of secretory activity by the epithelium of these organs, lipidic droplets and lamellar granules present in mature males' seminal vesicles and the vacuoles present in post-vesicular vas deferens are probably secretion. Besides, the spermatozoa in the lumen are immersed in a material of characteristic structure, which must be produced in superior regions of the reproductive system of immature males, not studied here. The presence of sperm cells, apparently in cytoplasm vesicles of seminal vesicle and post-vesicular vas deferens, suggests spermiophagy by their epithelium.

Effect of narcosis with CO2 on the ovarian development in queens of Apis mellifera (Hymenoptera, Apini)

Exposure of 1, 4, 7 & 10 day-old virgin queens of Apis mellifera Linne 1758 for 1 min to CO2 accelerated their ovarian development, having a similar effect as mating on the initial formation of the ovarian follicles. In 3 day-old queens the exposure to CO2 enhanced the initial stage of germ cell differentiation into oocytes and nurse cells in the ovarioles, a developmental stage only seen in 5 day-old untreated virgin queens, the age at which they are ready to mate. In 10 day-old untreated virgin queens, some regions of the ovarioles presented tissue disorganization and many cells with pycnotic nuclei. However, narcotized virgin queens of the same age did not present such ovary degeneration. Conversely, they showed nitid follicle formation, arising in the ovarioles' initial differentiation between nurse and oocytic chambers, although still without vitellogenesis. The accelerative effect of CO2 is limited to the ages near to those proper for mating, since 15 and 18 day-old treated virgin queens presented ovaries with extensive regions of high tissue disorganization and a great number of cells with pycnotic nuclei. According to the results, the narcosis presented three levels of effect on the ovary of honeybee virgin queens: 1) accelerated the germ cell differentiation, 2) preserved the tissue integrity even after the queen mating period and 3) stimulated the initial differentiation of a vitellarium. This later condition was only seen in untreated queens after mating. All these effects are not maintained if the queen is kept virgin beyond 15 days old.

Ultrastructural features of the regenerative cells of the bees (Hymenoptera, Apidae) midguts

The epithelium of the bee ventriculus is formed by two cell types: the principal or digestive cells and the regenerative cells. In this article the ultrastructure of the regenerative cells is described, as well as the features of their differentiation into digestive cells during epithelium renewal.

Comparative study of the ultrastructure and secretory dynamic of hypopharyngeal glands in queens, workers and males of Sceptotrigona postica latreille (Hymenoptera, Apinae, Meliponini)

The secretory cycle of hypopharyngeal glands (HPGs) in Scaptotrigona postica resembles that of Apis mellifera: in newly emerged workers the HPGs are in prefunctional state, their maximum development happens in the nurse workers and in forager workers they show signs of reabsorption. In S. postica these glands are also present in queens and males where they are more developed in newly emerged individuals. The ultrastructural features of the HPG secretory cycle in workers of S. postica and A. mellifera are alike: granular endoplasmic reticulum well developed, large secretion masses around the intracellular canaliculus in nurse workers and extensive degenerative structures in forager workers. Then it is suggested that the HPG secrete similar substances in both species. A second secretory cycle seems to occur in early foragers, may be with production of enzymes. The role of the HPGs in queens and males remains unknown but one possibility is enzyme production.

Differentiation of the worker's ovary in Apis mellifera L. (Hymenoptera, Apidae) during life of the larvae

The aim of the present study is to characterize the way worker and queen ovaries differentiate in, Apis mellifera, a species with trophic determination of female castes. A morphological study carried out with light and transmission electron microscopy showed that the differences in ovary development between the two castes begin as soon as the differential nursing of larvae is initiated. The decrease in ovariole number in worker ovaries is due to a process of cell death occurring in germinative cells and autophagic regression of somatic cells in the ovarioles that commence in the third instar larvae and proceed until the fifth instar where the process is more intense. Germinative cell death leads to ovariole disintegration and incorporation of the remaining somatic cells of the latter into the stromatic cells in such a way that the total volume of the ovary is little affected during larval development, although the ovariole number decreases. By the end of the larval stage, loss of cells is observed among the stromatic cells of the ovary. As a result, the ovary starts to decrease in volume and takes on the adult form.

Structural characterization of a new acylpolyaminetoxin from the venom of Brazilian garden spider Nephilengys cruentata

Mario Sergio Palma, Yasuhiro Itagaki, Tsuyoshi Fujita, Hideo Naoki and Terumi Nakajima. Structural characterization of a new acylpolpaminetoxin from the venom of Brazilian garden spider Nephilengys: cruentata. Toxicon 36, 455-493, 1998.-The use of mass spectrometry, in which high-energy CID and charge remote fragmentation both of protonated and sodium-attached molecular ions was applied, afforded the structural elucidation of a new acylgolyaminetoxin with M-W= 801 da from the venom of the Brazilian garden spider Nephilengys cruentata. In spite of having the same M-W of the NPTX-2, previously described in the venom of the Joro spider Nephila clavata, neither toxins are isomers. In order to differentiate them by using the most usual nomenclature, the new toxin was named NPTX-801C and the NPTX-2 was renamed to NPTX-801E. Both toxins have as common structure the 4-hydroxyindole-3-acetyl-asparaginyl-cadaveryl moiety in their molecules and their structure may be represented in a simplified way: NPTX-801E is HO-indole-Asn-Cad-Pta-Orn-Arg and NPTX-801C is HO-indole-Asn-Cad-Gly-Put-Pta-Pta. (C) 1998 Elsevier B.V. Ltd. All rights reserved.

Mass spectrometric characterization of two novel inflammatory peptides from the venom of the social wasp Polybia paulista

The social wasp P. paulista is relatively common in southeast Brazil causing many medically important stinging incidents. The seriousness of these incidents is dependent on the amount of venom inoculated by the wasps into the victims, and the characteristic envenomation symptoms are strongly dependent on the types of peptides present in the venom. In order to identify some of these naturally occurring peptides available in very low amounts, an analytical protocol was developed that uses a combination of reversed-phase and normal-phase high-performance liquid chromatography (HPLC) of wasp venom for peptide purification, with matrix-assisted laser desorption/ionization time-of-flight post-source decay mass spectrometry (MALDI-Tof-PSD-MS) and low-energy collision-induced dissociation (CID) in a quadrupole time-of-flight tandem mass spectrometry (QTof-MS/MS) instrument for peptide sequencing at the sub-picomole level. The distinction between Leu and Ile was achieved both by observing d-type fragment ions obtained under CID conditions and by comparison of retention times of the natural peptides and their synthetic counterparts (with different combinations of I and/or L at N- and C-terminal positions). To distinguish the isobaric residues K and Q, acetylation of peptides was followed by Q-Tof-MS analysis. The primary sequences obtained were INWLKLGKMVIDAL-NH2 (MW 1611.98Da) and IDWLKLGKMVMDVL-NH2 (MW 1658.98Da). Micro-scale bioassay protocols characterized both peptides as presenting potent hemolytic action, mast cell degranulation, and chemotaxis of poly-morphonucleated leukocyte (PMNL) cells. The primary sequences and the bioassay results suggest that these toxins constitute members of a new sub-class of mastoparan toxins, directly involved in the occurrence of inflammatory processes after wasp stinging. Copyright (C) 2004 John Wiley Sons, Ltd.

Morphological features and occurrence of degenerative characteristics in the hypopharyngeal glands of the paper wasp Polistes versicolor (Olivier) (Hymenoptera : Vespidae)

Different histochemical techniques were applied to examine the morphological features of the secretory cells of hypopharyngeal glands in the wasp Polistes versicolor. The results showed that most analyzed individuals present active glands with secretion stored in the cytoplasm. In some glands, morphological analyses revealed the presence of degenerative characteristics. Analyses of cellular integrity, however, did not detect dead cells. The results showed that, in P. versicolor, the development and regression of the hypopharyngeal glands were not age related, unlike glands of social bees. (c) 2006 Elsevier Ltd. All rights reserved.

Lipid distribution in salivary glands of larvae and adult bees (Hymenoptera : Apidae)

Cytochemical studies were carried out to establish lipid distribution in the salivary glands of larvae and adult bees, using the imidazole buffer technique. In the duct cells of the larval salivary gland, the reaction was positive in the epicuticle and negative in the glandular lumen. The absence of smooth endoplasmic reticulum and the presence of lipids in the intercellular space suggest that lipids absorbed from the haemolymph could be used in the constitution of the epicuticle, after having been conveyed through the epithelium. In adult workers (new-emerged, nurse and forager workers), the head salivary glands presented a positive reaction in the secretion in glandular lumen, identifying its lipidic nature.

Enzymatic activity of hypopharyngeal gland extractsfrom workers of Apis mellifera (Hymenoptera, Apidae, Apinae)

This research presents a comparative study of enzymatic activity of the hypopharyngeal gland extracts from workers of Apis mellifera in three physiologic stages: newly emerged, nurse and forager workers, with the objective of contributing to the comprehension of the gland function. In order to determinate the enzymes present in the extracts, the Api Zym kit (Bio Merieux) was used to test the activity of 19 different enzymes. The enzymes found in larger amounts only in the hypopharyngeal glands from certain individuals were the following: in newly emerged workers, the N-acetyl-double down arrow-glucosaminidase that may be digesting the chitin of some food ingested by the bee; in forager workers, the acid phosphatase that is likely acting in authophagic processes, the a-glucosidase, in the processing of nectar into honey, and the double down arrow-glucosidases, in the pollen digestion.

A new type of sternal gland present in Oxaea flavescens (Hymenoptera, Oxaeinae): location and histology

The sternal glands of the abdomen of Oxaea flavescens consist of class ill glandular cells, with the excretory canaliculus linking to the lateral intersegmental membrane of segments ill, TV and V. The intersegmental membrane is augmented and folded into several lobes forming a reservoir covered by secretory cells. The intersegmental membrane is then transformed into an intima that lines a reservoir space containing secretions of a type of mucus which is periodic acid-Schiff positive. The storage of a great amount of secretion suggests that it is not used continuously. These glands are absent from males, indicating that their products must have a specific function linked to the female sex.