External and internal structure of Atta bisphaerica Forel (Hymenoptera: Formicidae) nests

This work was carried out to determine the internal and external structures of Atta bisphaerica (Forel) nests. Six nests were excavated and during excavation, all data referring to chambers and tunnels were recorded. Three nests had been internally cement-moulded, which enabled a better view of the chamber and tunnel structures. Atta bisphaerica nests presented a similar structural pattern, varying only in the number of chambers as a function of external mound area. Chambers were spherical with two communication tunnels. Internal tunnels had an elliptical section, sometimes circular, indirectly linked to chambers through ramifications and directly through short tunnels. Entrance holes were linked to the area of highest chamber concentration by tunnels from the elliptical section, which led to the nest in a radial manner. Knowledge of the colony's three-dimensional architecture permits successful application of chemical control processes, reduces the quantity of product applied, and consequently diminishes costs and environmental damage.

The ant (Hymenoptera: Formicidae) community in two species of the Euphorbiaceae, Alchornea sidifolia and Croton floribundus, in an area of the Atlantic Forest of Brazil

The number of individuals and species richness of ants were investigated in two species of Euphorbiaceae found in three areas of differentiated flora. These areas form part of a fragment of the Atlantic Forest located in the Serra do Itapety, in the state of São Paulo, Brazil. Fifty eight ant species were sampled, belonging to 23 genera and 6 sub-families; 40% of the species are arboreal. The sub-family Myrmicinae was the richest in species, with Pheidole being the most diverse genus and Crematogaster the most commonly-occurring. In the three study areas, no significant differences were found between the number of individuals and the species richness of ants collected on the trunks of Euphorbiaceae. C. floribundus Spreng and A. sidifolia Muell. Arg. are similar with respect to the number of ant species sampled; the number collected being 40 and 47, respectively.

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.

The influence of fruit morphology and habitat structure on ant-seed interactions: A study with artificial fruits

Artificial fruits designed to simulate lipid-rich non-myrecochorous diaspores were used to test for the effect of fruit morphology and habitat structure on ant-seed interactions in an Atlantic Forest site in SE Brazil. The outcome of the interaction (i.e., if the fruit was removed, cleaned by ants on the spot or had no interaction with ants) and the time of ant response were the investigated variables. Models simulating drupes and arilate diaspores were used to test for morphological effects and four habitat attributes (litter depth, number of logs, number of trees, and percentage of bromeliad coverage on the forest floor), likely to be correlated with the ant diversity and abundance in the study site, were measured to test for the effect of habitat structure. The proportion of fruits removed or cleaned did not differ between the two morphological models. Sites in which fruits were cleaned had more trees than those in which no interaction occurred. This may be a result of the foraging behavior of arboreal ants that frequently descend to the forest floor to exploit fleshy diaspores. Sites in which model removal occurred had lower litter depth than both those in which models were cleaned and those in which no interaction occurred. A negative correlation was observed between litter depth and ant response time. Accumulation of leaf litter at a given point may have constrained the movements of large ants in general, and ponerine ants (that are important seed removers) in particular. We conclude that that local pattern in litter depth and tree density influence the frequency and outcome of interactions between ants and non-myrmecochorous, fleshy diaspores.

Macromolecular array patterns of silk gland secretion in social Hymenoptera larvae

The cocoon, produced by most holometabolous insects, is built with silk that is usually produced by the larval salivary gland. Although this silk has been widely studied in the Lepidoptera, its composition and macromolecular arrangement remains unknown in the Hymenoptera. The macromolecular array patterns of the silk in the larval salivary gland of some meliponids, wasps, and ants were analyzed with polarized-light microscopy, and they were compared with those of Bombyx mori (Lepidoptera). There is a birefringent secretion in the glandular lumen of all larvae, due to filamentous structural proteins that display anisotropy. The silk in the distal, middle and proximal regions of the secretory portion of Formicidae and Vespidae glands presented a lattice optical pattern. We found a different pattern in the middle secretory portion of the Meliponini, with a zigzag rather than a lattice pattern. This indicates that the biopolymer fibers begin their macromolecular reorganization at this glandular region, different from the Formicidae and the Vespidae, in which the zigzag optical pattern was only found at the lateral duct. Probably, the mechanism of silk production in the Hymenoptera is a characteristic inherited from a common ancestor of Vespoidea and Sphecoidea; the alterations in the pattern observed in the Meliponini could be a derived characteristic in the Hymenoptera. We found no similarity in the macromolecular reorganization patterns of the silk between the Hymenoptera species and the silkworm.

Toxicity of sesame extracts to leaf-cutting ant Atta sexdens rubropilosa (Hymenoptera: Formicidae)

Extracts of different sesame plant (Sesamum indicum, Linnaeus) organs were tested through ingestion and contact experiments to investigate their toxicity to Atta sexdens rubropilosa (Forel) workers. Dichloromethane extracts of seeds were toxic to the leaf-cutting ants and the factor responsible for the toxicity does not show seasonal occurrence.

Toxicity of sesame seed to leaf-cutting ant Atta sexdens rubropilosa (Hymenoptera: Formicidae)

Extracts of the ripe seeds of the sesame plant (Sesamum indicum, Linnaeus) were tested through contact experiments to investigate their toxicity to Atta sexdens rubropilosa workers. Dichloromethane extract of seeds was toxic to the ants and the factor responsible for this effect was distributed through the ethyl acetate fraction. This fraction was divided into four sub fractions composed of: A) triglycerides, B) monoglycerides + diglycerides + triglycerides, C) diglycerides + sesamoline + sesamine and D) sesamine. However, when these sub fractions were separated, no toxicity was observed. Therefore, in order to determine why the activity was lost, the concentration of each sub fraction was duplicated, and the possible combinations among them were also tested. We concluded that the toxicity to the ants is due mainly to a mixture of triglycerides, and sesamoline or the combination of sesamoline + sesamine can be a synergistic factor in this fraction.

Ocorrência de formigas Acromyrmex Mayr, 1865, em alguns municípios do Brasil

It were collected species of Acromyrmex Mayr, 1865 in many brazilian cities from Santa Catarina, Paraná, São Paulo, Minas Gerais, Goiás, Mato Grosso do Sul, Mato Grosso, Rondônia and Amapá states and in Paraguay and Uruguay, with the purpose to bring up-to-date their occurrence. Specimens collected on trips were identified at laboratory of Fca/Unesp/Botucatu-SP. It were recorded, for the first time, the occurrence of A. rugosus rugosus (F. Smith, 1858) in Paraná, Rondônia and Santa Catarina states; A. subterraneus subterraneus (Forel, 1893) in Amapá and Bahia; A. diasi Gonçalves, 1982 in Rio Grande do Sul and Paraná states; A. coronatus (Fabricius, 1804) in Paraná; A. balzani (Emery, 1890) in Amapá and A. subterranens brunneus (Forel, 1911) in Goiás.

Astilbin toxicity to leaf-cutting ant Atta sexdens rubropilosa (Hymenoptera: Formicidae)

The toxicity of astilbin, isolated from Dimorphandra mollis, was tested in laboratory ingestion bioassays for the leaf-cutting ant Atta sexdens rubropilosa Forel. Worker ants that were fed an artificial diet daily to which astilbin was added had a higher mortality rate than the controls. The substance astilbin was incorporated into the diet with and without the utilization of a solvent. Results for both methodologies utilized were similar and indicate that astilbin reduces the median survival of ants. All concentrations tested showed toxic effects against ant workers, and the statistical comparison of survivorship rates from control and treated groups was significantly different, confirming the insecticidal properties of the substance astilbin.

Comparative study of the fat body in some genera of the attini tribe (Hymenoptera: Formicidae)

The Cyphomyrmex rimosus Spinola and Mycetarotes parallelus Emery species of ants, considered basal, and Acromyrmex disciger Mayr and Atta laevigata Smith, considered derived, have fat bodies specially distributed on their gaster, around their organs and right below the cuticle. The fat body is formed by trophocytes, which are characterized by their pronounced vacuolization of the cytoplasm and the irregular morphology of their nuclei caused by the pressure exerted by cytoplasmic vacuoles. In C. rimosus, the nuclei are more regular, presenting an oval or a star form. In A. disciger and A. laevigata the nuclei present chromatin in a cord form, while in C. rimosus and M. paralellus the chromatin is uniformly distributed in the nucleoplasm, very condensed in the latter species. The parietal trophocytes of A. disciger show cytoplasm with a smaller quantity and smaller sizes of vacuoles compared to ones from the perivisceral region, the opposite is observed in C. rimosus. In A. laevigata and M. parallelus there were no differences observed in their cytoplasm between both regions of cells. In the trophocytes of C. rimosus, A. disciger, A. laevigata, there was a reticular aspect of the cytoplasm observed in the region between vacuoles, not seen on M. parallelus. Another cellular type, oenocyte, was found associated with the fat body cells, and is dispersed between trophocytes with an inner contact to them, but no membrane fusion with them. The oenocytes have a spherical form and are smaller than the trophocytes; they have acidophilic cytoplasm with a small quantity of small vacuoles, and round nuclei.

Toxicity of Cedrela fissilis to Atta sexdens rubropilosa (Hymenoptera: Formicidae) and its symbiotic fungus

Crude extracts from roots, stems, branches, fruits and leaves of Cedrela fissilis were tested to verify their toxicity to Atta sexdens rubropilosa workers and to their symbiotic fungus Leucoagaricus gongylophorus. The workers that were fed daily on an artificial diet to which crude extracts from this plant were added had a higher mortality rate than the controls, especially for the hexane, dichloromethane and methanol crude extracts from roots (RH, RD and RM) and from leaves (LH, LD and LM). Fungal growth was inhibited by the hexane (RH) and dichloromethane crude extract from roots (RD). The RH, RD and FD crude extracts were fractioned and their fractions were tested. All the fractions tested presented toxicity to the ants and some fractions (RH-H, RH-D, RD-4 and RD-5) completely inhibited fungus development. The possibility of controlling these insects in the future using Cedrela fissilis compounds that can simultaneously target both organisms is discussed.

Interference of epicuticular wax from leaves of grasses in selection and preparation of substrate for cultivation of symbiont fungus by Atta capiguara (Hym. Formicidae)

The selection of plants is one of the stages of foraging behavior, executed by leaf-cutting ants for the cultivation of symbiont fungus. Anatomical, biochemical and physiological characteristics of vegetal species are some of the factors that influence selection of substrate; however, most studies described in the literature refer to leaf-cutting ants. Due to the scarcity of studies on the role of epicuticular waxes of grasses in relation to foraging behavior of monocotyledonous leaf-cutters, we completed three experiments with the ant species Atta capiguara in which we analyzed: the attractiveness or selectivity of leaves of Saccharum officinarum (sugarcane) and/or Hypharrenia rufa (capim jaraguá), with and without epicuticular wax in relation to the worker of this ant species, their processing and elimination in the form of pellets of epicuticular wax from the referred fragments removed by the workers for cultivation of symbiont fungus. We observed that vegetal fragments without wax were those most transported by workers to the inside of the nests; that they processed the fragments equally with as much as without wax, and eliminated, in the form of pellets, wax removed from the same into garbage chambers of the colonies, leading us to conclude that for monocotyledonous leaf-cutters, epicuticular wax from vegetal leaves has great importance in vegetal integrity by being used as substrate, representing a physical barrier to the growth of symbiont fungus.

Study of the venom glands in Ectatomma quadridens (Hymenoptera: Formicidae). Evolutionary hypothesis in subfamily ponerinae

The venom glands of worker ants of the species Ectatomma quadridens morphologically resemble an elongated sac or reservoir ending in a narrower portion that has the function of releasing the secretion to the exterior. Two external secretory filaments are individually inserted into the proximal portion of the gland and end inside the convoluted gland. The venom gland of workers of E. quadridens is, therefore, morphologically subdivided into four distinct portions: a) sac-shaped reservoir measuring approximately 1mm in length; b) excretory duct, proximal portion of the reservoir that joins the gland to the sting apparatus; c) convoluted gland, final portion of the external secretory filaments located inside the reservoir; and d) two secretory filaments measuring about 2 mm in length; their free extremities end blindly and are individually inserted into the reservoir wall at the proximal region of the venom gland. The histological data showed that the filaments and the convoluted gland are composed of cubic cells of secretory function. The reservoir consists of a simple cubical epithelium externally surrounded by muscle fibers. A thick cuticle internally coats the epithelium of the reservoir. The application of histochemical tests allowed us to establish that the final secretion of the venom gland of Ectatomma quadridens is of glycoproteic nature. This secretion undergoes several modifications at the secretory filaments, at the convoluted gland, and in the reservoir before reaching the excretory duct, the point at which the secretion is released in its final composition, namely the venom. Based on the differences among various Ponerinae species we propose a hypothesis suggesting a probable evolutionary process that the venom glands of members of this subfamily might have undergone.

A new imidazole alkaloid and other constituents from Pilocarpus grandiflorus and their antifungal activity

The stems of Pilocarpus grandiflorus have afforded the new imidazole alkaloid 4,6-dehydro-1,2,4,5-tetrahydro-2,5-dioxopilocarpine in addition to the 17 known compounds germanicol, β-amiryn, ocotillone, stigmast-4-en-3-one, 3β-hydroxy-stigmast-5-en-7-one, 6β-hydroxy-stigmast-4-en-3-one, β-sitosterol, scopoletin, 3-(1′,1′-dimethylallyl)-scopoletin, elisin, dictamine, 4-methoxy-2-quinolone, platydesmine, syringaresinol, syringaldehyde, syringic acid and vanillic acid. Their structures were elucidated on the basis of chemical and spectroscopic evidence. The phenolic compounds vanillic acid and syringaldehyde and the furoquinoline alkaloid platydesmine exhibited antifungal activity against Leucoagaricus gongylophorus, the symbiotic fungus of leaf-cutting ants (Atta sexdens rubropilosa). © 2005 Verlag der Zeitschrift für Naturforschung.