Fungal communities in the garden chamber soils of leaf-cutting ants

Leaf-cutting ants modify the properties of the soil adjacent to their nests. Here, we examined whether such an ant-altered environment impacts the belowground fungal communities. Fungal diversity and community structure of soil from the fungus garden chambers of Atta sexdens rubropilosa and Atta bisphaerica, two widespread leaf-cutting ants in Brazil, were determined and compared with non-nest soils. Culture-dependent methods revealed similar species richness but different community compositions between both types of soils. Penicillium janthinellum and Trichoderma spirale were the prevalent isolates in fungus chamber soils and non-nest soils, respectively. In contrast to cultivation methods, analyses of clone libraries based on the internal transcribed spacer (ITS) region indicated that richness of operational taxonomic units significantly differed between soils of the fungus chamber and non-nest soils. FastUnifrac analyses based on ITS sequences further revealed a clear distinction in the community structure between both types of soils. Plectania milleri and an uncultured Clavariaceae fungus were prevalent in fungus chamber soils and non-nest soils, respectively. FastUnifrac analyses also revealed that fungal community structures of soil from the garden chambers markedly differed among ant species. Our findings suggest that leaf-cutting ants affect fungal communities in the soil from the fungus chamber in comparison to non-nest soils. © 2013 WILEY-VCH Verlag GmbH & Co.

Association between Pseudonocardia symbionts and Atta leaf-cutting ants suggested by improved isolation methods

Fungus-growing ants associate with multiple symbiotic microbes, including Actinobacteria for production of antibiotics. The best studied of these bacteria are within the genus Pseudonocardia, which in most fungus-growing ants are conspicuously visible on the external cuticle of workers. However, given that fungus-growing ants in the genus Atta do not carry visible Actinobacteria on their cuticle, it is unclear if this genus engages in the symbiosis with Pseudonocardia. Here we explore whether improving culturing techniques can allow for successful isolation of Pseudonocardia from Atta cephalotes leaf-cutting ants. We obtained Pseudonocardia from 9 of 11 isolation method/colony component combinations from all 5 colonies intensively sampled. The most efficient technique was bead-beating workers in phosphate buffer solution, then plating the suspension on carboxymethylcellulose medium. Placing these strains in a fungus-growing ant-associated Pseudonocardia phylogeny revealed that while some strains grouped with clades of Pseudonocardia associated with other genera of fungus-growing ants, a large portion of the isolates fell into two novel phylogenetic clades previously not identified from this ant-microbe symbiosis. Our findings suggest that Pseudonocardia may be associated with Atta fungus-growing ants, potentially internalized, and that localizing the symbiont and exploring its role is necessary to shed further light on the association.

Energy substrate used by workers of leaf-cutting ants during nest excavation

Energy substrate used by workers of leaf-cutting ants during nest excavation. In this study we aimed to ascertain whether leaf-cutting ant workers lose body reserves (fat or sugars) as a function of nest excavation. For each treatment, we isolated 10 workers of Atta sexdens into two experimental groups, Control (C- without excavation) and Soil (S- with excavation), which were kept for different time intervals (0, 24, 48 or 72 hours), totaling 700 tested workers. We then determined the concentration of soluble carbohydrates and total lipid content in them. The total carbohydrates were determined colorimetrically, based on the reaction between carbohydrates and sulfuric acid-phenol. For determination of lipids, the insects were immersed in organic solvent until they reached a constant weight. Our results showed that carbohydrates are consumed during nest excavation activities. In the experimental groups S24, S48 and S72, there was an average reduction of 5.82 (20.42%), 14.31 (44.96%) and 13.27 (43.96%) µ.mg-1 in soluble sugar when compared with the experimental groups that did not excavate. Furthermore, the lipids were not used during this activity. With respect to dry mass of the workers, their values were C0 = 8%, C24 = 10.4%, C48 = 9.2%, C72 = 10%, S24 = 9.2%, S48 = 8.7% and S72 = 8.5%. Our results show experimentally that the source of energy for nest excavation is carbohydrates, whereas lipids are conserved for other activities.

Novel Phialophora species from leaf-cutting ants (tribe Attini)

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Leaf-cutting ants: an unexpected microenvironment holding human opportunistic black fungi

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Mitochondrial and peroxisomal population in post-pharyngeal glands of leaf-cutting ants after lipid supplementation

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

What is the stimulus for the excavation of fungus chamber in leaf-cutting ants?

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Comparative study of resistance to heat in two species of leaf-cutting ants

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Acromyrmex charruanus: a new inquiline social parasite species of leaf-cutting ants

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

First-year nest growth in the leaf-cutting ants Atta bisphaerica and Atta sexdens rubropilosa

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

The influence of temperature on oxygen consumption rates of workers of two leaf cutting ants, Atta laevigata (F. Smith, 1858) and Atta sexdens rubropilosa (Forel, 1908)

1. 1. The oxygen consumption in workers of two simpatric leaf cutting ants, Atta laevigata and Atta sexdens rubropilosa was measured at different temperatures. 2. 2. In the temperature range between 5-35°C, with 5°C increments, the respiratory rates increased with temperature, but the R-T curves of both ants showed neither a marked drop at the low end nor a break at the high end; except between 30 and 35°C. 3. 3. The respiratory rates of A. s. rubropilosa were higher than those of A. laevigata and in the midrange of temperatures, the rates of A. laevigata increased faster than those of A. s. rubropilosa. 4. 4. Q10 values did not indicate regions of compensation for temperature in both ants, but suggested that adjustments may occur at high temperatures (25-35°C), as expected for tropical ants. 5. 5. Temperature variations did not alter significantly the slope of the curve relating oxygen consumption and body weight in both species. © 1982.

The respiratory metabolism of the castes of two leaf cutting ants, Atta laevigata (F. Smith, 1858) and Atta sexdens rubropilosa forel, 1908

1. 1. The respiration of the different castes of two species of leaf-cutting ants, Atta laevigata and Atta sexdens rubropilosa was measured in both workers and sexed forms. 2. 2. The respiratory rates (μl O2/mg(live)/hr) were, in A. laevigata, 0.644 (gardeners), 0.594 (cutters), 0.354 (soldiers), 0.233 (males) and 0.085 (females); in A. s. rubropilosa, 0.803 (gardeners), 0.748 (cutters), 0.510 (soldiers), 0.375 (males) and 0.274 (females), significantly higher than A. laevigata. 3. 3. All castes considered, the b-values of the equation relating oxygen consumption and weight in A. laevigata (0.53) were significantly different from that in A. s. rubropilosa (0.73). © 1981.

Shared Escovopsis parasites between leaf-cutting and non-leaf-cutting ants in the higher attine fungus-growing ant symbiosis

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Considerations for Assessing Maximum Critical Temperatures in Small Ectothermic Animals: Insights from Leaf-Cutting Ants

The thermal limits of individual animals were originally proposed as a link between animal physiology and thermal ecology. Although this link is valid in theory, the evaluation of physiological tolerances involves some problems that are the focus of this study. One rationale was that heating rates shall influence upper critical limits, so that ecological thermal limits need to consider experimental heating rates. In addition, if thermal limits are not surpassed in experiments, subsequent tests of the same individual should yield similar results or produce evidence of hardening. Finally, several non-controlled variables such as time under experimental conditions and procedures may affect results. To analyze these issues we conducted an integrative study of upper critical temperatures in a single species, the ant Atta sexdens rubropiosa, an animal model providing large numbers of individuals of diverse sizes but similar genetic makeup. Our specific aims were to test the 1) influence of heating rates in the experimental evaluation of upper critical temperature, 2) assumptions of absence of physical damage and reproducibility, and 3) sources of variance often overlooked in the thermal-limits literature; and 4) to introduce some experimental approaches that may help researchers to separate physiological and methodological issues. The upper thermal limits were influenced by both heating rates and body mass. In the latter case, the effect was physiological rather than methodological. The critical temperature decreased during subsequent tests performed on the same individual ants, even one week after the initial test. Accordingly, upper thermal limits may have been overestimated by our (and typical) protocols. Heating rates, body mass, procedures independent of temperature and other variables may affect the estimation of upper critical temperatures. Therefore, based on our data, we offer suggestions to enhance the quality of measurements, and offer recommendations to authors aiming to compile and analyze databases from the literature.

Influence of caffeine on the survival of leaf-cutting ants Atta sexdens rubropilosa and in vitro growth of their mutualistic fungus

BACKGROUND: Leaf-cutting ants collect plant fresh material for the cultivation of their mutualistic fungus. Atta sexdens rubropilosa Forel (Hymenoptera: Formicidae) cause great economic losses through their foraging activity, mainly in agriculture. The main control method is the application of granulated toxic baits incorporated with an active ingredient (AI). The present goal is to evaluate the effect of caffeine on in vitro growth of the mutualistic fungus and on the survival of the leaf-cutting ants, aiming to verify the potential toxicity of this secondary metabolite over these organisms. RESULTS: Three distinct patterns of fungal growth correlated with caffeine concentration were observed: (1) no effect (0.01% caffeine); (2) intermediate growth reduction (0.05% caffeine); (3) drastic growth reduction (0.10 and 0.50% caffeine). The highest caffeine concentration causes fungus death in the first week. As for insect survival, caffeine does not seem to exert any effect. The treatments with diet containing caffeine showed similar values of M50, irrespective of caffeine concentration. CONCLUSION: As caffeine was shown to reduce growth of the mutualistic fungus of Atta sexdens rubropilosa, but with no conclusive effect on insect survival, a hypothetical explanation for the selection of different Coffea species by this leaf-cutting ant species might be associated with caffeine toxicity to the fungus. Copyright (C) 2011 Society of Chemical Industry