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.

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.

Note about the first swarming in a laboratory colony of Coptotermes gestroi (Isoptera: Rhinotermitidae)

A laboratory rearing of Coptotermes gestroi which swarmed from the reared colony is described.

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.

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.

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.

Structure of the spermatheca in five families of isoptera

The structure of the spermatheca was investigated in specimens of five termite families with the aid of light microscopy. In longitudinal section, the spermatheca of Zootermopsis nevadensis (Termopsidae) showed the shape of an umbrella with a secretory portion and duct. The other termite species, which belong to the families Kalotermitidae, Serritermitidae, Rhinotermitidae and Termitidae showed a spermatheca constituted only by the secretory portion. This structure was an elongate, fingerlike tube with a recurved and blind extremity. The spermatheca wall was composed of a single epithelium formed by class 3 secretory cells with a lumen lined by cuticle. The cuticle was thin and smooth or thick with digitiform projections in the species examined. All the termite females showed bundles of musculature outside of the spermatheca.

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.

The caste system of Coptotermes gestroi (Isoptera: Rhinotermitidae)

Coptotermes gestroi is an oriental species introduced to Brazil and considered, in the São Paulo State area, one of the most economically important pests. Although there are a few works concerning the basic biology of this species, its post-embryonic development has been poorly studied. The aim of the present research was to study the post-embryonic development of C. gestroi for a better knowledge of the caste system in this termite. The post-embryonic development of C. gestroi starts with two larval instars of whitish appearance and different sizes. A separation of the neutral and imaginal developmental pathways occurs following the second larval instar. The second-instar larva originates alates after six molts. The worker caste presents five instars and also develops from the second larval instar. Soldiers develop from younger workers in laboratory colonies and from older workers in field colonies.

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.

Ant-tended hemiptera in Amazonian myrmecophytes: Patterns of abundance and implications for mutualism function (Hymenoptera: Formicidae)

We assessed how the abundance of ant-tended Hemiptera associated with two Amazonian myrmecophytes, Tococa bullifera and Maieta guianensis, varied as a function of resident ant species. We collected five species or morpho-species of adult hemiptera in the domatia of M. guianensis, with four of these species also found in Tococa bullifera. Maieta guianensis plants inhabited by Crematogaster laevis had over four-fold more hemiptera in them than plants inhabited by Pheidole minutula. In contrast, the density of hemiptera in Tococa bullifera domatia was independent of the species of ant resident. For each of the two ant species inhabiting Maieta guianensis, there was a positive and significant relationship between the abundance of hemiptera and workers inhabiting a plant. This relationship was also significant and positive for the Tococa bullifera plants inhabited by C. laevis. However, there was no relationship between Azteca worker and hemipteran density, although there was a trend towards a positive relationship. Our results indicate that hemipteran abundance can vary significantly between different myrmecophyte species, but that the nature of this relationship is mediated by the identity of the ant associate. Because hemipterans are herbivores, the costs and benefits of different ant partners to the host plant may vary in ways that are often overlooked.

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.