The effect of plant diversity on fungus garden development and foraging behavior of leaf-cutting ants (Hymenoptera: Formicidae)

The leaf-cutting ants forage a wide variety of plant species, used for symbiotic fungus cultivation. To better understand this tripartite complex interaction, 24 colonies of Acromyrmex subterraneus brunneus were conditioned for 4 months to 6 different plants (Citrus spp., Ligustrum spp., Acalypha spp., Eucalyptus spp., Alchornea triplinervia, Melia spp.), to verify the influence of conditioning on foraging behavior of workers. The effect of plants on symbiotic fungus development was studied separately, through macerated plants in Agar and culture medium A as the control. During foraging, workers presented polyphagic foraging behavior, refusing the plants to which they were conditioned. The selection of plants is not correlated with the plant substrate that promotes good development of symbiotic fungus. Such results demonstrate the importance of plant diversity for fungus garden maintenance.

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.

Carbon isotope composition and leaf anatomy as a tool to characterize the photosynthetic mechanism of Artemisia annua L.

Leaves of Artemisia annua L. are a plentiful source of artemisinin, a drug with proven effectiveness against malaria. The aim of this study was to classify the photosynthetic mechanism of A. annua through studies of the carbon isotope composition (δ 13C) and the leaf anatomy. A. annua presented a δ 13C value of - 31.76 ± 0.07, which characterizes the plants as a typical species of the C3 photosynthethic mechanism, considering that the average δ 13C values for C3 and C4 species are -28 and -14, respectively. The leaf anatomy studies were consistent with the δ 13C results, where, in spite of the existence of parenchymatic cells forming a sheath surrounding the vascular tissue, the cells do not contain chloroplasts or starch. This characteristic is clearly different from that of the Kranz anatomy found in C4 species.

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.

Inhibition of the symbiotic fungus of leaf-cutting ants by coumarins

Leaf-cutting ants are known to be a serious pest for agriculture due to the high amounts of vegetal matter from crops used by them in order to cultivate a symbiotic fungus on which they rely for food and enzymes. The mutualism between the fungus and the ants is a point to be explored when alternative methods of control are being thought of. Considering that some plants are naturally resistant to phytophagous insects, some natural products (secondary metabolites) should be evaluated with respect to their insecticide and/or fungicide properties. In this paper we isolated eight coumarins from four different plant species and we determined their effect on the development of the symbiotic fungus of the leaf-cutting ant Atta sexdens. With the exception of clausarin, all the other coumarins were inhibitory from 64 μg mL-1 through 80 μg mL-1 and xanthyletin inhibited the fungus at 25 μ mL -1. ©2005 Sociedade Brasileira de Química.

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.

Leaf-cutting ants toxicity of limonexic acid and degraded limonoids from Raulinoa echinata. X-ray structure of epoxy-fraxinellone

Phytochemical survey of roots extracts of the South Brazilian endemic plant Raulinoa echinata Cowan, Rutaceae led to the isolation of known degraded limonoids: fraxinellone, fraxinellonone and epoxy-fraxinellone. The latter was previously isolated also from the stems, but the relative configuration could not be solved by NMR experiments. This paper deals with the X-ray diffraction analysis of epoxy-fraxinellone and its toxicity to leaf-cutting ants, describes the isolation of fraxinellonone and fraxinellone for the first time in Raulinoa. Epoxy-fraxinellone showed no toxicity to the leaf-cutting ants (Atta sexdens rubropilosa). The limonoid limonexic acid, isolated from stems, presented high toxicity to the leaf-cutting ants, diminishing considerably their longevity. ©2005 Sociedade Brasileira de Química.

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.

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.

Toxicity of straight-chain fatty acids to leaf-cutting ants Atta sexdens rubropilosa (Hymenoptera: Formicidae) and the symbiotic fungus Leucoagaricus gongylophorus

This work determined toxicity and attractiveness of straight-chain fatty acids (C 5 to C 12) to Atta sexdens rubropilosa (Forel) workers. The effect to the symbiotic fungus, Leucoagaricus gongylophorus (Singer) Möller, was also tested with the fatty acids C 6 to C 12. A strong mortality of leaf-cutting ants that were fed with an artificial diet containing fatty acids C to C at concentrations above 1.0 mg.ml -1 was observed. Rice flakes impregnated with solutions of these fatty acids were repellent to leaf-cutting ants. Contact experiments showed that treatments with C 6 and C 7 at concentration of 100 mg.ml -1 significantly reduced the survival rate of leaf-cutting ants. The fatty acids C 8 to C 11 were toxic to leaf-cutting ants when topically tested at concentration of 200 mg.ml -1. In relation to the fungus' bioassays, the fatty acids C 6 to C 12 at concentration of 0.1 mg.ml -1 inhibited 100% of the fungal development. Although when the concentration was reduced by half no inhibition effects were observed. The results showed that straight-chain fatty acids have desirable properties for controlling leaf-cutting ants since they directly interfere with both organisms of the symbiotic relationship. The potential of fatty acids as well as ways to control leaf-cutting ants with these compounds are discussed in this article.

The role of workers in transferring queen substances and the differences between worker castes in the leaf-cutting ant, Acromyrmex subterraneus brunneus (Hymenoptera: Formicidae)

The present study focused on the relationship between the queen and workers in Acromyrmex subterraneus brunneus colonies mediated by the possible transfer of substances from the fertile to the sterile caste. The queens were submitted to different situations of physical limitation, i.e., they were kept isolated in cages with holes that only permitted the entry of workers but left the queen isolated. A tracer (water-soluble dye) was applied to the pronotum and gaster of the queen and its dispersal among workers was analyzed. The results demonstrated that the minor sub caste (0.7-0.9 mm) passed on the dye through allogrooming and self-grooming, or transferred the dye through trophallaxis to the major sub caste (1.2-2.0 mm) when they were not in direct contact with the queen. These findings indicate the communication and probable transfer of substances from the queen to the workers, as well as a substance transfer between workers.

Evaluation of antidiarrhoeal effects of Psidium guajava L. (Myrtaceae) aqueous leaf extract in mice

A crude aqueous extract of the leaves of the guava tree, Psidium guajava L. (Myrtaceae), were studied for antidiarrhoeal effects, to place one of its traditional medical uses. The extract induced a decrease in the propulsive movements of the intestinal contents in mice. These findings suggested that an aqueous extract of guava leaves may be used as an effective treatment for non-specific diarrhoea in folk medicine.

Dispersal of the delayed action insecticide sulfluramid in colonies of the leaf-cutting ant Atta sexdens rubropilosa (Hymenoptera: Formicidae)

Toxic baits are the most used control method for leaf-cutting ants due to their high effectiveness and because they are considered the safest for humans. Taking into account that the importance of leaf-cutting ants as pests, knowing the process by which dispersal and worker contamination is achieved becomes essential to understand several aspects about the functioning of a bait-borne AI (active ingredient) used in toxic baits. It has been established that an effective toxic bait should have a delayed- action AI, but its dispersion among the different sizes of workers is unknown. Workers of different sizes are involved in quite different tasks such foraging, cultivation of symbiotic macrofungus or control of deleterious microfungi. Therefore, we prepared a toxic bait containing the delayed-action AI sulfluramid and a dye (Rhodamine B) as an AI tracer in order to study dispersal and contamination in colonies, evaluated at different periods and in relation to different workers' sizes. Both field and laboratory colonies were evaluated. The great level of contamination, about 50% at 24 hours, in all sizes of workers demonstrates that worker contact with toxic bait is intense within this period. The distribution in field and laboratory colonies was similar. This contamination pattern is probably enough to cause the colony to die because of contamination of smaller workers, leading to the loss of control of the aggressive microfungi, which can quickly overgrow the symbiotic fungus culture. The dispersal dynamics of AI in leaf-cutting ant workers is important for investigations on the mode of action of this insecticide in the colony, and as a reference in future studies, such as those attempting to reduce the concentration of AIs in baits to reduce their environmental impact, or for facilitation of new AI screening.