RAPD analysis of the sexual state and sterile mycelium of the fungus cultivated by the leaf-cutting ant Acromyrmex hispidus fallax

That the symbiotic fungus of leaf-cutting ants only occasionally produces the sexual phase makes their identification confusing. This has occurred so rarely, either in laboratory nests, or in unbalanced field nests. that the possibility of contamination of the fungal garden by other fungi cannot be disregarded. In this paper we describe the formation of several basidiomata in a healthy and free-living nest of the leaf-cutting ant Acromyrmex hispidus fallax. The cultivation in vitro of the sterile mycelia (isolated from the fungal garden) with their typical inflated tips, and the similarity of both forms confirmed by RAPD analysis of their genomic DNA. The fungus was identified as Leucoagaricus gongylophorus.

Age polyethism in the leaf-cutting ant Acromyrmex subterraneus brunneus Forel, 1911 (Hym., Formicidae)

This study describes and quantifies the behavioural acts of two laboratory colonies of Acromyrmex subterraneus brunneus by investigating worker age polyethism. Twenty-nine behavioural acts were recorded during the 19-week observation period. Young individuals performed tasks inside the nest related to brood care and care for the fungus garden, whereas older individuals performed activities outside the nest such as foraging and activities in the waste chamber. The average longevity (+/- SD) was 108.21 +/- 3.30, 109.15 +/- 1.92 and 122.71 +/- 1.55 days for large, medium and small workers, respectively. The small-sized workers presented a higher probability of reaching older age than large- and medium-sized workers. This study describes task switching according to age polyethism and the relationship of physical and temporal subcastes.

THE POPULATION STATUS OF THE ENDANGERED BRAZILIAN ENDEMIC LEAF-CUTTING ANT ATTA-ROBUSTA (HYMENOPTERA, FORMICIDAE)

The population status of the endemic leaf-cutting ant Atta robusta was assessed using extensive field surveys. In the coastal plain of the state of Rio de Janeiro, we found localized populations of A. robusta, bur no evidence of population recruitment. In disturbed habitats, A. robusta is apparently being displaced by Atta sexdens rubropilosa.

Yeasts associated with nests of the leaf-cutting ant Atta sexdens rubropilosa Forel, 1908

A total of 137 yeasts associated with the leaf-cutting ant Atta sexdens rubropilosa Forel, 1908 were characterized, being selected 93 for analysis. Twenty four species belonging to seven genera(Candida, Cryptococcus, Rhodotorula, Sporobolomyces, Tremella, Trichosporon, Pichia) were isolated from the different analysed material. The genus Candida was widely distributed, with C. homilentoma, C. colliculosa-like, C. famata and C. colliculosa being the most prevalent. A few isolates did not fit the standard descriptions and probably some of them could be new biotypes or even new species. Three strains of black yeasts were also isolated, and four others were identified as being Candida spp. The effective number of yeast species was higher in newer sponge. The origin, distribution and relative importance of these microorganisms for the ants are discussed.

GROWTH OF OOCYTE AND NURSE CHAMBER DURING OOGENESIS IN QUEENS OF THE LEAF-CUTTING ANT ATTA SEXDENS RUBROPILOSA FOREL, 1908 (HYM, FORMICIDAE)

Volume changes of the vitellarium components of the leaf-cutting ant Atta sexdens rubropilosa are reported. The oocyte grew to approximately 409 times of its initial volume and reached a maximum value of 1.2 x 10(7) mu m(3). The follicle increase in size at a more or less constant rate up to the 12(th), showing an elevated growth rate thereafter. The mean number of follicles per ovariole was 16.42 +/- 3.58.

Sex allocation in the polydomous leaf-cutting ant Acromyrmex balzani

Logistic regression analysis was used to analyse sex allocation in a population of the leaf-cutting ant Acromyrmex balzani occurring in a pasture in southern Brazil. The field sample consisted of 151 fungus-garden chambers (18 queenright and 133 queenless), belonging to 50 nests with three vertically stacked chambers per nest on average. Taking nest chamber as the unit of analysis, seven predictor variables were considered: sampling date, chamber depth, chamber volume, weight of fungus garden, presence of a queen, number of large workers, and number of small to medium workers. The population-level numerical proportion of females was 0.548 and the inferred proportional energetic investment in females 0.672. The former was not significantly different from 0.5 (P=0.168), but the latter was (P=0.0003). The proportional investment in females per fungus garden increased with the number of large workers present (P=0.0002) and decreased with the dry weight of the fungus garden (P=0.012). This implies that resource acquisition through foraging is likely to be a major proximate determinant of sex allocation. The negative correlation between female bias and fungus garden weight might be due to developing adult females requiring more food than males, but this hypothesis could not be confirmed by direct statistical evidence.

Cellulose degradation by Leucocoprinus gongylophorus, the fungus cultured by the leaf-cutting ant Atta sexdens rubropilosa

Leucocoprinus gongylophorus, the fungus cultured by the leaf-cutting ant Atta sexdens rubropilosa, is able to degrade efficiently cellulose, microcrystaline cellulose, carboximethylcellulose, and cellobiose. Analysis of the degradation products indicate that the fungus produce extracellular β-glucosidase, exo- and endo-glucanase. The importance of cellulose degradation to the association of fungus and ant is discussed.

Effect of constituents of the foliar wax of Didymopanax vinosum on the foraging activity of the leaf-cutting ant Atta sexdens rubropilosa

Leaf epicuticular waxes may affect substrate selection by leaf-cutting ants, and host recognition by several phytophagous insects. The influence of the crude epicuticular wax of Didymopanax vinosum E. March. (Araliaceae), and its major constituents lupeol and free primary alcohols, on substrate selection by the leaf-cutting ant Atta sexdens rubropilosa Forel, was investigated. Two ant colonies were used in laboratory bioassays. Corn flakes (zea) were impregnated with chloroform solutions of the substances being tested and fed to the adults. One of the colonies gave results, in most of the tests, not significantly different from the controls. The other colony, however, discriminated against the crude wax, a strong deterrent effect being observed from the first of a series of eight trials. The same colony was able to discriminate against lupeol after the second trial. The fraction of primary n-alcohols (22% C28, 66% C30 and 12% C32) deterred feeding only after the fifth trial. The results demonstrate that some constituents of epicuticular waxes may deter the foraging activity of A. sexdens rubropilosa, depending, however, on the colony under observation. It is suggested that lupeol, of the foliar wax of D. vinosum, is an important deterrent to leaf-cutting ants, although with less pronounced effects than those observed in tests with the crude wax.

Occurrence of killer yeasts in leaf-cutting ant nests

Killer activity was screened in 99 yeast strains isolated from the nests of the leaf-cutting ant Atta sexdens against 6 standard sensitive strains, as well as against each other. Among this yeast community killer activity was widespread since 77 strains (78 %) were able to kill or inhibit the growth of at least one standard strain or nest strain. Toxin production was observed in representatives of all the studied genera including Aureobasidium, Rhodotorula, Tremella and Trichosporon, whose killer activity has not yet been described.

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.

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 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.

Queen lipid content and nest growth in the leaf cutting ant (Atta sexdens rubropilosa) (Hymenoptera: Formicidae)

The relationship between the queens' lipid content and nest growth (population size, biomass and nest architecture) was studied from founding up to 1 year. Nests aged 3, 4, 5, 6, 9 and 12 months were dug in the field, and their dimensions were measured. The ant nest population and fungus garden was also collected. The sample was taken to the laboratory where we counted the worker population and weighed the biomass (fungus plus offspring) and queens. Queens were separated for the determination of lipids. The lipid content in the bodies of queens decreased in the first months, then stabilized (at 4-6 months) before increasing in months 9 and 12. Nest biomass (symbiotic fungus and offspring) and worker population increased over time. The structural growth of the nests was observed by excavating around them. Initially nests (3 months old) had one chamber at an average depth of 15 cm. By 1 year, the nests had three or four deep chambers, and were about 3-4 m deep. Our study contributes to knowledge of the dynamics of the energy-reserve expenditure by queens during colony founding and colony development for up to 1 year. © 2013 Copyright Taylor and Francis Group, LLC.