Ecological interaction between Atta sexdens (Hymenoptera: Formicidae) and the vegetation of a mesophyll semideciduous forest fragment in Botucatu, SP, Brazil

Among 89 plants species growing in a subtropical dry forest fragment located in Botucatu, State of São Paulo, Brazil, 35 species were cut by Atta sexdens, representing 39.34% of the current plants existing in this area. A. sexdens L., 1758 (Hymenoptera: Formicidae) has a selective preference for the following species: Alchornea triplinervia, Faramea cyanea, Cariniana estrellensis and Casearea obliqua, with the first being the most cut species. The frequency and absolute densities of the plant families and species significantly influence the selection process. When comparing the absolute frequency regarding the 10 most cut plant species and the cutting frequency, significant data were obtained only for the euphorbiaceous A. triplinervia species, proving the preference of A. sexdens for this species in particular. These interactions are discussed based on ecological and management factors in agro-ecosystems.

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.

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.

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.

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.

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.

Toxicity of Cipadessa fruticosa to the leaf-cutting ants Atta sexdens rubropilosa (Hymenoptera: Formicidae) and their symbiotic fungus

Crude extracts from fruits, branches and leaves of Cipadessa fruticosa were assayed on Atta sexdens rubropilosa workers and their symbiotic fungus Leucoagaricus gongylophorus. The results showed that the majority of the extract tested reduced significantly (p < 0.05) the survival of the workers comparable to the neem oil. In addition, the dichloromethane extract from fruits and the hexane extract from branches of C. fruticosa inhibited in 80% the development of the symbiotic fungus. The hexane and dichloromethane extracts from fruits of C. fruticosa were fractionated and their fractions were subjected to new bioassays. Several fractions showed relevant activity on ants, but none of them inhibited significantly the fungal growth. The possibility of controlling leaf-cutting ants in the future using C. fruticosa is discussed.

Assessment of microfungi in fungus gardens free of the leaf-cutting ant Atta sexdens rubropilosa (Hymenoptera: Formicidae)

The purpose of this study was to determine the microfungi present in young nests of Atta sexdens rubropilosa when the fungus gardens were deprived of worker ants. The results were compared with another study in which worker ants had been killed by using toxic baits, and some species such as Acremonium kiliense, Escovopsis weberi, Moniliella suaveolens and Trichoderma sp. were confirmed among the most common inhabitants of this microenvironment, but differences in composition and proportion of species were observed. The importance and the role of these and other species of fungi within the symbiosis are discussed.

The hypopharyngeal gland of leaf-cutting ants (Atta sexdens rubropilosa) (Hymenoptera: Formicidae)

The social behavior of ants is controlled by means of dynamic interactions between the environment and the individuals, mainly through pheromones. Among the exocrine glands is the hypopharyngeal gland (HG), located laterally over the pharyngeal plate. The present work aimed to describe and compare the HG in the different castes of the leaf-cutting ant Atta sexdens rubropilosa Forel 1908. The HG of different castes showed similar morphology varying only in relation to the secretory cell number. The secretory intracellular reservoir presented positive reaction to Xylidine Ponceau and P.A.S, indicating the presence of protein and polysaccharides in the secretion. Therefore, we suggest that the function of this gland would be the production of digestive enzymes and/or some sort of mucus, which, together with the secretions produced by the salivary glands of the thorax, would be related to the production of saliva.

Attractiveness of citrus pulp and orange albedo extracts to Atta sexdens rubropilosa (Hymenoptera: Formicidae)

The present study aimed to compare the attractiveness of industrial citrus pulp with the handmade orange albedo to the workers of Atta sexdens rubropilosa. For this, filter paper fragments were impregnated with organic extracts obtained through chemical extraction and sequential fractioning with hexane and dichloromethane and offered to different field nests. It was verified that the industrial citrus pulp extract is as good as the handmade orange albedo extract. This preference is discussed keeping in mind the chemical, behavioral and nutritional factors.

Morphohistochemical characterization of the perivisceral fat body in royal and worker female castes in different ages of Atta sexdens rubropilosa ants (Hymenoptera, Formicidae)

The morphohistochemical study of the fat body in royal and worker castes of Atta sexdens rubropilosa revealed that this tissue is made up of trophocytes and oenocytes, trophocytes being the cells with cytoplasmic inclusions and irregular nuclei and the oenocytes cells with homogeneous cytoplasm and rounded nuclei. The trophocytes of queens presented many lipidic and proteic granules, while those of workers presented trophocytes with fine granulation in the cytoplasm constituted mainly by lipids and carbohydrates. The oenocytes from the worker caste of A. s. rubropilosa presented cytoplasm filled mainly with lipoproteic material.

Toxicity of Azadirachta indica to leaf-cutting ant Atta sexdens rubropilosa (Hymenoptera: Formicidae)

The leaf-cutting ants are responsible for great economical damages in cultivated areas, and because of this characteristic, they have aroused the interest of researchers around the world. Thus, extracts of neem (Azadirachta indica, A. Juss) were tested through ingestion and contact experiments to investigate their toxicity to Atta sexdens rubropilosa workers. Crude extract of neem seed oil was toxic to the ants by ingestion in all concentrations tested but no toxicity was observed in contact experiments. However, crude extract of neem seed paste produced a significant toxicity to workers by ingestion and by contact.