Enzymes present in the thoracic gland extracts from workers and males of Apis mellifera (Hymenoptera: Apidae)

This research deals with the analysis of the enzymes present in thoracic gland extracts from newly emerged, nurse workers, forager workers, newly emerged males, and mature males of A. mellifera L. (Hymenoptera, Apoidea, Apidae). The enzymes found in larger quantities in the thoracic gland occurred in all classes of workers and are digestive. Acid phosphatase and Naphtol-AS-BI-phosphohydrolase act in protein synthesis, leucine arylamidase hydrolyses proteins and a-glucosidase actuate in the nectar processing into honey. Naphtol-AS-BI-phosphohydrolase was found in larger quantities only in workers, this suggests action in protein synthesis by the thoracic gland, b-galactosidase is in larger amounts in the newly emerged bees (workers and males) this aids in the provision of other substances to be used as an energy source when glucose or sucrose are absent. Differences between enzymatic profiles from workers and males are usually related to their colony tasks, or related to their physiological necessities per individual in specific life stages.

Behavior of Atta sexdens rubropilosa (Hymenoptera: Formicidae) workers during the preparation of the leaf substrate for symbiont fungus culture

The behavioral repertory of Atta sexdens rubropilosa Forel (Hymenoptera: Formicidae) workers marked by size category was studied during the preparation of the leaf substrate in the laboratory. The workers were marked according to three physical castes, i.e., minima, generalist and forager. Seven types of behavioral acts were recorded for each caste, with the following frequencies: licking leaf fragments (64.6%), holding fragments on the surface of the fungus garden (16.4%), shredding the fragments (6.0%), chewing and crimping the edges of the fragments (9.0%), incorporating the fragments (2.7%), touching the surface of the fungus with their mandibles and other mouthparts after incorporation (0.3%), and depositing fecal fluid (0.1%). The minima workers were found to be more specialized in the activities related to the preparation of the leaf substrate, which represented 52% of the total number of tasks performed. The generalists performed 40.3% of these tasks, and the foragers 7.9%. Licking the substrate was the behavior most frequently recorded and performed for a longer period of time. In this way, the workers may feed and at the same time eliminate microorganisms that are harmful to the symbiont fungus. The smaller castes, minima and generalists, are those most responsible for the preparation of the leaf substrate and predominate within a colony. From a practical viewpoint, with the introduction of toxic bait containing insecticides, for example, these size categories will be those most intensely intoxicated, especially through the behavior of licking bait pellets. On the basis of the data obtained about these behaviors, we may raise the hypothesis that trophallaxis in not the major factor triggering contamination with an insecticide among the workers of a colony.

Enzymatic activity of hypopharyngeal gland extracts from 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 Mérieux) 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-down double arrow sign-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 down double arrow sign-glucosidases, in the pollen digestion.

Enzymes in the hypopharyngeal gland extracts from workers of Scaptotrigona postica (Hymenoptera, Apinae, Meliponini) related to food storing in the colony

This study reports on research of enzymes produced by the hypopharyngeal glands, which are related to food storing in the colony, from gland extracts from nurse and forager workers of S. postica. Only the presence of the saccharase was detected in the extracts from the glands of forager workers. The results were compared to the enzymatic content of similar extracts of A. mellifera taking into account the behavioral differences among the two species.

Enzymatic activity of the hypopharyngeal gland extract from workers and males of Scaptotrigona postica Lat. (Hymenoptera, Apidae, Meliponini)

The objective of this research was to contribute to elucidation of the function of the hypopharyngeal glands of S. postica in enzyme production, using the Api Zym kit (Bio Mérieux). Dealing with a comparative analysis between the enzymatic content of the hypopharyngeal gland extracts from newly emerged, nurse and forager workers, as well as, newly emerged males and males mature for mating of S. postica. The hypopharyngeal glands from nurse workers of Apis mellifera, that produce part of the royal jelly, were used for comparison. While in A. mellifera, the hypopharyngeal glands are present only in workers, in S. postica, the hypopharyngeal glands are present and functioning in all adult individuals of the colony. The higher enzymatic activity was observed in the hypopharyngeal gland extracts from nurse workers and may be related to a larger demand for energy, compared to other individuals. The occurrence of large quantities of leucine arylamidase in all individuals may mean that protein processing is happening.

Temporal polyethism in workers of Ectatomma edentatum (Formicidae: Ponerinae)

In the present study, adult ants emerged from five nests kept under laboratory conditions were individually monitored in order to determine their longevity and observe their individual behavior in relation to age (temporal polyethism). Our results showed that most of the tasks related to colony maintenance were executed during the three first age intervals (comprising from 1 to 150 days of age), namely brood care, followed by grooming activities (self and allogrooming), nest maintenance activities (such as floor and nest inspection), and social related activities (antennating and food exchange among adults). Foraging activities were performed by ants in the higher age intervals.

Alimentary preconditioning in Acromyrmex subterraneus brunneus workers

Ants suffer a strong environmental influence and are sometimes conditioned for many social activities. To understand better this phenomenon in leaf-cutting ants, 24 colonies of Acromyrmex subterraneus brunneus (Hymenoptera: Formicidae) were conditioned for 2 months with 6 different plants (Citrus spp., Ligustrum spp., Acalypha spp., Eucalyptus spp., Alchornea triplinervia, Melia spp.), to verify alimentary preconditioning in plant selection. After the conditioning period, plants were offered simultaneously, to evaluate the acceptability of each plant species. During foraging, the workers did not exhibit alimentary source fidelity, which was familiarized. This fact suggests the absence of alimentary preconditioning, reinforcing the polyphagic foraging behavior of leaf-cutting ants.

Size of fat body trophocytes and the ovarian development in workers and queens of Melipona quadrifasciata anthidioides

The correlation between trophocyte size and ovarian development is negative in workers and positive in queens of Melipona quadrifasciata anthidioides. The nurse workers which have the ovaries in a higher developmental stage, present smaller fat body cells them newly-emerged ones. In newly-emerged and nurse workers the trophocytes seem to be delivering their stored products, among which probably vitellogenin. As in workers the cell size variations do not support the occurrence of proteic synthesis or the increasing in reverses storage after the adult emergence, the products released from the trophocytes must come from cellular reserves remaining from the larval phase. This datum is in agreement with the early and brief vitellogenic phase in the ovaries of this caste. In foragers the trophocyte size seen stabilized. In queens it was verified considerable increasing in the trophocyte size from virgin to physogastric queen, as well as the maintenance of the size during all fertile life of the queen.

Comparative study of the cuticular hydrocarbon composition of Melipona bicolor Lepeletier, 1836 (Hymenoptera, Meliponini) workers and queens

In social insects, cuticular hydrocarbons are involved in species, kin, caste and nestmate recognition. Gas chromatography and mass spectrometry were used to compare the cuticular hydrocarbon composition of workers, males and queens of Melipona bicolor. The cuticular hydrocarbon composition of this species was found to consist mainly of C23, C25:1, C25, C27:1, C27, C29:1 and C29, which are already present in imagoes that have not yet abandoned the brood cell. This composition varied quantitatively and qualitatively between and within the castes and sexes. The newly emerged workers and young queens (virgins) had similar cuticular hydrocarbon profiles, which were different from those of the males. When the females start executing their tasks in the colony, the cuticular hydrocarbon profile differences appear. The workers have less variety, while the queens conserve or increase the number of cuticular hydrocarbon compounds. The queens have more abdominal tegumentary glands than the workers, which apparently are the source of the new cuticular compounds.

Apis mellifera (Hymenoptera, Apini) Ovary Development in Queens and in Workers from Queenright and Queenless Colonies

A morphological study of the development of the ovaries in workers from queenright and queenless colonies and queens of Apis mellifera is presented and discussed. The mating stimulates the ovary development and the egg-laying. Mating prevention or egg laying prevention provokes reabsorption in the queens ovaries, characteristic of the queen stage or ovariole development. Ovary development was observed in queenless and in queenright colonies but the queen's presence, the worker's age and the type of tasks done by them interfere with their development.

Juvenile Hormone Effect on the Venom Gland Secretory Cycle in Workers of Apis mellifera (Hymenoptera, Apidae)

The present investigation analyzed the influence of Juvenile Hormone (JH) on the venom glands of Apis mellifera workers through protein dosage and electrophoresis of venom gland extracts of newly emerged workers which were treated with 1 μl JH dissolved in hexane, in concentration of 2μg/μl. Newly emerged workers non-treated and treated with 1 μl hexane were the controls. Both JH and hexane provoke quantitative changes on the gland protein titre and on the protein electrophoretic profile. The disappearance of protein bands in the venom gland extracts of 14 day-old treated workers, a situation normally found only in 35 day-old non-treated workers, suggests that the JH treatment induces a precocious maturation of the worker venom gland.

Histochemistry and protein profile of the mandibular glands of workers of the ant Atta sexdens rubropilosa (Hymenoptera: Formicidae)

Insect mandibular glands are always associated to the mandibles; they are part of the salivary glandular system. The mandibular glands are composed by a reservoir associated to the secretory cells, with each secretory cell connected to the reservoir by means of individual canaliculi. These glands play an important role in the production of pheromones, which are compounds involved in defense, communication, and reproduction of the colony. Mandibular glands of soldiers and major and minor workers of the ant Atta sexdens rubropilosa were processed for different histochemical tests, total protein content, and protein electrophoretic profile determination. The histochemical tests detected the presence of lipids, DNA/RNA, polysaccharides, and proteins at different regions of the gland. The protein electrophoretic profiles showed that the total protein content as well as the number of peptides of each caste follow a progressive order in relation to the size of the individual. Thus, we suggest that the production of secretion is directly linked to the task that the individual performs in the colony.

Method for the evaluation of insecticidal activity over time in Atta sexdens rubropilosa workers (Hymenoptera: Formicidae)

The active ingredients used in the formulation of toxic baits for leaf-cutting ants (Hymenoptera: Formicidae) should possess a delayed action defined as an insecticidal activity whereby worker mortality is ≤15% at 24 hours and ≥90% at 21 days. Serious shortcomings have occurred in the search for new active ingredients, such as the initial selection of fenoxycarb, copper oxychloride and diflubenzuron, compounds considered very promising but whose inefficiency was verified only later, indicating methodological problems. In view of this situation, we developed a classification method for insecticidal activity over time using workers of the leaf-cutting ant Atta sexdens rubropilosa Forel. The insecticides used were fipronil, sulfluramid GX071HB and sulfluramid GX439, vehicled in an attractive pasty formulation prepared based on citrus pulp. The results obtained were consistent from a toxicological viewpoint and agreed with the literature in terms of the control of colonies. Sulfluramids were found to possess a delayed action at a broad range of concentrations, in agreement with the fact that these substances are highly effective in the control of all leaf-cutting ant species. The smaller range of concentrations of fipronil with delayed action is probably related to its lower efficacy for species more difficult to control such as Atta capiguara (Forti et al. 2003). We discuss the importance of relating behavioral particularities to the specific feeding habits of leaf-cutting ants, with methodological adequacy of the assessment of insecticides aimed at toxic baits.

Temporal polyethism and survivorship of workers of Agelaia pallipes (Hymenoptera, Vespidae, Epiponini)

In this study, known-age workers of Agelaia pallipes (Olivier) were introduced into a colony maintained in the laboratory and the effects of age polyethism on the worker life span were observed. As in other Neotropical swarm-founding wasps, the results showed that younger workers remained on different places inside the nest performing activities related to the brood and nest maintenance, while risky tasks such as foraging were performed by older workers. Furthermore, the start of foraging activities had a significant effect on worker mortality.

Ultramorphology and histology of the venom gland of workers of Apis mellifera (Hymenoptera, Apidae) before and after an electrical shock treatment

This paper describes the ultramorphology and histology of the venom reservoir in 14-day old workers of Apis mellifera, immediately before and after the application of electrical shocks with the object of causing venom elimination and reservoir collapse. The external epithelial surface of the reservoir was differentiated according to its morphological aspects into posterior, median, and proximal or duct regions at the ventral surface and into anterior and posterior regions at the dorsal surface. While the epithelium of the proximal region forms a ventral infolding, a dorsal salience is formed at this region. These structures and the epithelial regions persist both in full and empty reservoirs. The reservoir appeared full and distended before the electrical shocks were applied and became empty and withered afterwards due to the elimination of the secretion, without any reductions in length. Nevertheless, some secretion was kept inside the lumen, thus suggesting a possible role for the reservoir in the modification of the secretion.