The grass-cutting ant Atta bisphaerica is one of the most serious pests in several pastures and crops in Brazil. Fungal diseases are a constant threat to these large societies composed of millions of closely related individuals. We investigated the occurrence of filamentous fungi associated with the ant A. bisphaerica in a pasture area of Viçosa, Minas Gerais State, Brazil. Several fungi species were isolated from forager ants, and two of them, known as entomopathogenic, Beauveria bassiana and Aspergillus ochraceus, were tested against worker ants in the laboratory. The two species were highly virulent, achieving 50 percent worker mortality within 4-5 days. It is the first time A. ochraceus, a commonly found fungal species, is reported to infect Atta species at a high prevalence. Possible uses for the fungus within biological control are discussed.
Acromyrmex leaf-cutting ants use the antibiotics produced by ectosymbionts (Actinobacteria) to suppress the growth of Escovopsis spp., which are the specialized parasites of attine fungus gardens. However, the spectrum of activity of these bacteria and their interactions with insect immune systems have not been thoroughly studied. The objective of this study was to evaluate whether the presence of symbiotic actinobacteria covering the entire cuticle of Acromyrmex subterraneus subterraneus (Forel) (Hymenoptera: Formicidae) workers provides protection against two fungus species, Metarhizium anisopliae (Metschn.) Sorokin (Hypocreales), a persistent entomopathogenic, and Aspergillus ochraceus G. Wilh. (Eurotiales), an opportunistic insect pathogen. We assessed the mortality rates of three groups of Ac. subterraneus subterraneus workers inoculated with a suspension of one of the two pathogens: external workers (EXT), internal workers with actinomycetes covering the whole body (INB), and internal workers without actinomycetes covering the whole body (INØ). We also assessed the encapsulation response, which is a parameter of innate immunity, to determine whether the pathogens affect immune responses in the ant. All bioassays were conducted on workers from seven laboratory colonies of Ac. subterraneus subterraneus. Mortality tests showed that actinomycetes do protect the workers of group INB against M. anisopliae, but not against A. ochraceus. Regarding the encapsulation response, INB ants treated with A. ochraceus conidia exhibited immunosuppression due to this pathogen. All three groups of workers that received M. anisopliae conidia presented some degree of immunosuppression after pathogen exposure. These results indicate that workers without visible bacteria cover, specially the external workers (EXT), are more resistant to entomopathogenic fungi. Nevertheless, bacterial actinomycetes present in young workers are important in protecting them from ubiquitous entomopathogenic fungi.
Founder females of the leaf-cutting ant species Atta sexdens experience high mortality during the founding and establishment of their colonies. The foundation site is crucial for the success of a new colony. In this study, we isolated and identified actinobacteria from fungus garden chambers of A. sexdens colony growth in soils from (1) forested areas without leafcutter ant nests and (2) open ground areas close to leafcutter ant nests. The inhibitory effect of these isolates on pathogenic fungi and the mutualistic fungus cultivated by leafcutter ants was evaluated. The 16S rRNA gene sequences were employed to identify nine selected actinobacteria species found in the soil: Streptomyces (6), Nocardia (2), and Kitasatospora (1). One Streptomyces and one Kitasatospora isolate inhibited all the tested fungi. Since there is no evidence of actinobacteria cultivation in the workers' cuticle of the Atta genus, our results corroborate the hypothesis that these workers may establish temporary adaptive symbiosis with soil microorganisms that produce antibiotic substances, living in some parts of their nest, or even inside their bodies.
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