The role of fungus in the diet of the leaf-cutting ant Atta cephalotes (L.)

Abstract. 1. The workers and queen of the leaf-cutting ant Atta cephalotes fed on the juice of swollen hyphae (staphylae) produced by their cultivated fungus, but neither obtained sufficient energy from this source for their respiratory needs. The number of staphylae eaten by workers increased with worker size but was not enough to satisfy their energy requirements.
2. Larvae fed on whole staphylae and staphylae previously chewed by workers, and obtained sufficient energy from this source for respiration and growth. No evidence of feeding on fungus hyphae or of trophallaxis between worker and larvae was found. Larvae preferred staphylae to hyphae when fed them artificially and they gained more weight on the former.
3. Worker ants imbibed plant sap during the preparation of plant material for the fungus garden and the uptake of carbohydrate during this process was sufficient to supply their energy needs for approximately 24 h.
4. Staphylae were richer in lipid and carbohydrate, and poorer in protein than ant fungal hyphae.
5. The number of staphylae produced by the fungus gardens of two small nests was comparable with the observed consumption rate but would provide only about 4% of the nest's respiratory requirements.
6. In the light of these findings, a revised view of the role of the fungus in the diet of the ant is discussed.     

Fungal hyphae as a source of nutrients for the leaf-cutting ant Atta sexdens

Abstract. The mutualistic fungus of leaf-cutting ants produces both ordinary hyphae and specialized ant rewards: the staphylae. Workers of Atta sexdens (L.) lived longer on diets which included nutritive staphylae than on those which provided only hyphae. However, hyphae were a better diet than sucrose solution or water alone. Small workers lived longer than large workers when receiving fungus garden, whether or not staphylae were available, probably because they are specialized to care for the garden and can exploit it. Large workers lived longer than small ones when only water was available. This may be a consequence of scale, larger workers containing proportionately more nutritional reserves in their bodies.
When starved workers were exposed to fungus for 3h, they gained weight. This weight gain represented the amount of food ingested during the test period. Workers gained 7.3 times more weight on natural fungus garden bearing staphylae than on garden with only hyphae. They gained only 1.5 times as much weight when the fungus was artificially cultured on agar. The results suggested that workers found hyphae attractive, but difficult to obtain in natural fungus gardens.
Material from hyphae and staphylae stained with a fluorescent dye was detected in worker crop contents using fluorescence microscopy. The crop contents of workers fed on staphylae fluoresced 1.14 times more than those of workers fed on hyphae.
Hyphae may provide a small source of food for workers, and the fungus as a whole may provide up to 9.0% of the respiratory energy requirements of workers, the remainder presumably being provided by plant sap.     

The role of substrate preparation in the symbiosis between the leaf-cutting ant Acromyrmex octospinosus (Reich) and its food fungus

Abstract. 1. In laboratory nests the ants thoroughly licked leaf fragments by a rasping action of their glossa before inoculating the food fungus. The extent of this licking varied with the substrate and was influenced by the thickness of surface leaf waxes and by the chemical attractiveness of the substrate.
2. Microscopy and wettability studies showed that licking removes the wax layer present on leaves. It also removes or inhibits micro-organisms present.
3. Chemically de-waxed and decontaminated leaves and ant-licked leaves proved the best substrates for artificially culturing the ant fungus.
4. The process of substrate preparation circumvents some of the normal defence mechanisms of green plants against fungal attack. The ant fungus thus utilizes nutritionally richer resources of green leaves normally available only to specialized parasitic fungi, rather than the nutritionally inferior shed leaves normally available to saprophytic fungi.     

The effects of leaf deprivation on leaf-cutting ants and their mutualistic fungus

1. When leaf-cutting ants were deprived of leaves for 5 days, they increased their consumption rates of 'staphylae', the nutritive bodies produced by their mutualistic fungus. They also pruned their fungus garden more intensively, presumably stimulating increased staphyla production.
2. Depriving fungus gardens of leaves for 5 days led to greater rates of staphyla production compared with control gardens. The rate of substrate exhaustion also increased, possibly because of this increased staphyla production, which led to greater removal of fungal resources by the ants.
3. The availability of leaf substrate directly affected staphyla production by the fungus garden. When leaf fragments were inserted into fungus garden samples, staphyla production was depressed, suggesting that the fungus can allocate resources either to hyphal colonization of fresh substrate or to staphyla production.
4. Under conditions of leaf deprivation, food production for the ants effectively increased, due to the combined effects of increased pruning by the ants and the response of the fungus to lack of substrate. This will allow nests to survive for long periods in the field when forage availability is reduced.
5. Exposing parts of the colony to the stress of leaf deprivation showed that there is a high level of flexibility in the fungus garden's ability to produce staphylae. This may be important during the cyclic production of broods, when demands for larval food will fluctuate widely. Under normal conditions, the optimal strategy may be for the fungus garden to have a moderate rate of production with a low rate of turnover, unless demand for staphylae increases dramatically.     

The origin of the attine ant-fungus mutualism.

Cultivation of fungus for food originated about 45-65 million years ago in the ancestor of fungus-growing ants (Formicidae, tribe Attini), representing an evolutionary transition from the life of a hunter-gatherer of arthropod prey, nectar, and other plant juices, to the life of a farmer subsisting on cultivated fungi. Seven hypotheses have been suggested for the origin of attine fungiculture, each differing with respect to the substrate used by the ancestral attine ants for fungal cultivation. Phylogenetic information on the cultivated fungi, in conjunction with information on the nesting biology of extant attine ants and their presumed closest relatives, reveal that the attine ancestors probably did not encounter their cultivars-to-be in seed stores (von Ihering 1894), in rotting wood (Forel 1902), as mycorrhizae (Garling 1979), on arthropod corpses (von Ihering 1894) or ant faeces in nest middens (Wheeler 1907). Rather, the attine ant-fungus mutualism probably arose from adventitious interactions with fungi that grew on walls of nests built in leaf litter (Emery 1899), or from a system of fungal myrmecochory in which specialized fungi relied on ants for dispersal (Bailey 1920) and in which the ants fortuitously vectored these fungi from parent to offspring nests prior to a true fungicultural stage. Reliance on fungi as a dominant food source has evolved only twice in ants: first in the attine ants, and second in some ant species in the solenopsidine genus Megalomyrmex that either coexist as trophic parasites in gardens of attine hosts or aggressively usurp gardens from them. All other known ant-fungus associations are either adventitious or have nonnutritional functions (e.g., strengthening of carton-walls in ant nests). There exist no unambiguous reports of facultative mycophagy in ants, but such trophic ant-fungus interactions would most likely occur underground or in leaf litter and thus escape easy observation. Indirect evidence of fungivory can be deduced from contents of the ant alimentary canal and particularly from the contents of the infrabuccal pocket, a pharyngeal device that filters out solids before liquids pass into the intestine. Infrabuccal pocket contents reveal that ants routinely ingest fungal spores and hyphal material. Infrabuccal contents are eventually expelled as a pellet on nest middens or away from the nest by foragers, suggesting that the pellet provides fungi with a means for the dispersal of spores and hyphae. Associations between such "buccophilous" fungi and ants may have originated multiple times and may have become elaborated and externalized in the case of the attine ant-fungus mutualism. Thus, contrary to the traditional model in which attine fungi are viewed as passive symbionts that happened to come under ant control, this alternative model of a myrmecochorous origin of the attine mutualism attributes an important role to evolutionary modifications of the fungi that preceded the ant transition from hunter-gatherer to fungus farmer.     

The influence of ants and water availability on oviposition behaviour and survivorship of a facultatively ant-tended herbivore

1. The lycaenid butterfly Hemiargus isola associates facultatively with the ant species Formica perpilosa in arid areas of south-western North America. Ants solicit liquid food rewards from butterfly larvae as larvae feed on the host plant, Acacia constricta . Previous studies have shown that tending by F. perpilosa enhances larval growth and pupal survivorship.
2. The effects of ants and plant water content on oviposition behaviour and survivorship to the last larval instar were tested by excluding ants and supplementing water to host plants in a two-way factorial experiment.
3. Butterflies, which lay eggs singly on host plant inflorescences, laid significantly higher egg numbers and densities (eggs/inflorescence) on plants with ants than on plants without ants. This is the first report of a facultative, generalized ant-associate using ants as oviposition cues. Water supplements increased the number, but not the density, of eggs laid on plants. Therefore, it appears that egg-laying butterflies responded to number of inflorescences, rather than plant tissue water per se .
4. Plants with ants had significantly greater numbers of inflorescences during the experiment than plants without ants. Water supplements increased number of inflorescences slightly, but not significantly.
5. Ants increased larval survivorship. Twice as many fourth-instar larvae survived per egg laid on plants with ants than on plants without ants. Ants did not reduce the number of predators present on acacias, but may have reduced predator effectiveness. Ants also did not reduce the numbers of potential H. isola competitors present.
6. Water supplementation affected neither the survivorship of H. isola larvae, nor the intensity of ant tending. Water supplementation did not affect the abundance of predators on plants, but did increase the abundance of several herbivorous insect taxa.     

Plant-ants feed their host plant, but above all a fungal symbiont to recycle nitrogen

In ant-plant symbioses, plants provide symbiotic ants with food and specialized nesting cavities (called domatia). In many ant-plant symbioses, a fungal patch grows within each domatium. The symbiotic nature of the fungal association has been shown in the ant-plant Leonardoxa africana and its protective mutualist ant Petalomyrmex phylax. To decipher trophic fluxes among the three partners, food enriched in (13)C and (15)N was given to the ants and tracked in the different parts of the symbiosis up to 660 days later. The plant received a small, but significant, amount of nitrogen from the ants. However, the ants fed more intensively the fungus. The pattern of isotope enrichment in the system indicated an ant behaviour that functions specifically to feed the fungus. After 660 days, the introduced nitrogen was still present in the system and homogeneously distributed among ant, plant and fungal compartments, indicating efficient recycling within the symbiosis. Another experiment showed that the plant surface absorbed nutrients (in the form of simple molecules) whether or not it is coated by fungus. Our study provides arguments for a mutualistic status of the fungal associate and a framework for investigating the previously unsuspected complexity of food webs in ant-plant mutualisms.     

The role of leaf-cutting ant workers (Hymenoptera: Formicidae) in fungus garden maintenance

Abstract.

• 1 We studied the role of leaf-cutting ant workers (Atta sexdens (L.) in fungus garden maintenance, by temporarily excluding workers from the garden. This increased its subsequent attractiveness, as expressed by an increase in the numbers of workers licking it.
• 2 The length of free mycelia on areas of the garden from which workers were excluded increased but was reduced again when workers were returned. Workers therefore removed hyphae from the garden surface.
• 3 The maximum‘isolation effect’was obtained by preventing ant access for 2–3 days, after which the effect declined. Removing staphylae from portions of garden kept ant-free for 4 and 6 days restored the effect, as the ants were not distracted by harvesting staphylae. Portions of garden kept ant-free for longer than this were no more attractive than non-isolated control garden.
• 4 Workers were highly efficient in detecting and removing contaminants from their fungus garden. Samples of garden could be isolated from workers for up to 12 days before major growth of contaminants occurred, and this contrasted with the maximum of 6 days for the isolation effect on licking. The isolation effect was therefore not a response to contaminant growths on the garden.
• 5 Workers on the garden surface may remove hyphae for nutritional reasons, or to‘prune’their fungus and stimulate its growth. In either case, the result is a regulation of fungal growth.

     

Larvae of Maculinea rebeli, a large-blue butterfly, and their Myrmica host ants: wild adoption and behaviour in ant-nests

The behavioural interactions between caterpillars of Maculinea rebeli Hir. and their Myrmica ant hosts were studied, both in the wild at the time of adoption, and inside captive nests of six Myrmica species.
In the wild, freshly moulted, final instar caterpillars left their food-plants at a time of day that coincided with the peak foraging activity of Myrmica (18:00-20:00 h). Once on the ground, caterpillars made no attempt to search for Myrmica but settled and waited for foraging ants to find them, which took up to 1.5 h. There was no adoption ritual: foragers of any Myrmica species picked up the caterpillars within 1–4 sec of discovery, and carried them directly to their nests.
Caterpillars grew from < 2 mg to 110 mg in laboratory ant-nests. About 60 mg was gained in autumn but 40% of this was lost during the winter, while the temperature was < 14 °C. Although caterpillars survived best with their normal host, Myrmica schencki , they could also survive in the nests of other Myrmica species. The presence of queen ants had no effect upon survival. The behaviour of the caterpillars was described and illustrated: this included the production of secretions that were drunk by the ants, begging for food and direct feeding by ants. The preferred solid food was ant eggs.
The results are discussed in terms of the social biology of Myrmica ants. It is hypothesized that Maculinea rebeli caterpillars mimic the touch pheromones of ant worker-larvae. This would explain the inability of ants to recognize caterpillars before touching them, their immediate adoption by any Myrmica species after discovery, host specificity inside wild ant-nests, the absence of queen-effect and the intimate attention of host workers.     

Floral visitation patterns of two invasive ant species and their effects on other hymenopteran visitors

Abstract.  1. Floral nectar of the native Hawaiian 'ōhi'a tree, Metrosideros polymorpha , is an important food source for several native honeycreepers and yellow-faced bees, Hylaeus spp., but is also attractive to invasive ants.
2. I undertook this study to compare floral visitation patterns of two widespread invasive ants, the Argentine ant, Linepithema humile , and the big-headed ant, Pheidole megacephala , and to determine their effects on nectar volume and floral hymenopteran visitors.
3. In the first year of the study, Argentine ants visited inflorescences more frequently than big-headed ants at mid-day and in the afternoon, but did not occur in higher densities than big-headed ants at any time of day. In the following year, Argentine ants visited inflorescences both more frequently and in higher densities than big-headed ants. Argentine ant density had a stronger association with nectar concentration than big-headed ant density.
4. Nectar volume did not differ between ant-excluded and ant-visited inflorescences for either ant species. However, ant density was negatively associated with nectar volume for both species.
5.  Hylaeus spp. never visited inflorescences with big-headed ants, while non-native honeybees visited inflorescences with and without ants of either species in equal frequency.
6. Most studies of the effects of invasive ants on native arthropods have focused on interactions on the ground. Flowers should not be overlooked as microhabitats from which native arthropods may be displaced by invasive ants.     

High Diversity and Low Specificity of Chaetothyrialean Fungi in Carton Galleries in a Neotropical Ant–Plant Association

New associations have recently been discovered between arboreal ants that live on myrmecophytic plants, and different groups of fungi. Most of the – usually undescribed – fungi cultured by the ants belong to the order Chaetothyriales (Ascomycetes). Chaetothyriales occur in the nesting spaces provided by the host plant, and form a major part of the cardboard-like material produced by the ants for constructing nests and runway galleries. Until now, the fungi have been considered specific to each ant species. We focus on the three-way association between the plant Tetrathylacium macrophyllum (Salicaceae), the ant Azteca brevis (Formicidae: Dolichoderinae) and various chaetothyrialean fungi. Azteca brevis builds extensive runway galleries along branches of T. macrophyllum. The carton of the gallery walls consists of masticated plant material densely pervaded by chaetothyrialean hyphae. In order to characterise the specificity of the ant–fungus association, fungi from the runway galleries of 19 ant colonies were grown as pure cultures and analyzed using partial SSU, complete ITS, 5.8S and partial LSU rDNA sequences. This gave 128 different fungal genotypes, 78% of which were clustered into three monophyletic groups. The most common fungus (either genotype or approximate species-level OTU) was found in the runway galleries of 63% of the investigated ant colonies. This indicates that there can be a dominant fungus but, in general, a wider guild of chaetothyrialean fungi share the same ant mutualist in Azteca brevis.     

Isolation, Growth Characteristics, and Long-Term Storage of Fungi Cultivated by Attine Ants

Seven pure-culture strains of fungi cultivated by attine ants (ant-garden fungi) were isolated from locally maintained leaf-cutting ant colonies. An ant-garden fungus strain obtained from an Atta cephalotes colony, when offered to ants of the colony from which the fungus was isolated, was accepted as their own. Young fungus cultures were harvested and incorporated into the fungus garden, and cultures of intermediate age were used to begin a new fungus garden; old cultures were simply harvested. To facilitate further research on this fungus, growth characteristics of the different isolates were studied under a variety of conditions. They grew better at 24°C than at 30°C, and growth did not occur at an incubation temperature of 37°C. In a broth culture medium, growth was enhanced by aeration of the culture and by addition of yeast extract, olive oil, sesame oil, peanut oil, soybean oil, corn oil, sunflower oil, cottonseed oil, walnut oil, safflower oil, or mineral oil. Glycerol did not noticeably affect growth, but Tween 80 inhibited growth. These fungi were extremely sensitive to cycloheximide, growth being totally inhibited at cycloheximide concentrations ranging from 0.4 to 4.0 μg/ml. To date, the ant-garden fungus isolates have remained viable in long-term mineral oil-overlay storage cultures for up to 4 years.     

Effects of ants on the foraging of birds in spruce trees

I experimentally excluded ants from randomly selected spruce trees Picea abies near colonies of the wood ant Formica aquilonia. Foraging activity of birds in these trees was then compared to the foraging activity of birds in neighboring spruce trees, where ants were allowed to continue foraging. Birds which foraged in the foliage showed the effects of competition with ants: they visited the trees without ants more frequently, and for longer periods. In addition, the insects and spiders that they utilized as food were more abundant in the foliage of trees without ants. Cone-foraging birds, however, which fed on seeds in cones at the tops of the trees, did not show a preference for trees without ants. The differences of tree usage between foliage-gleaning and coneforaging birds can be explained by alteration of the birds' food supply by wood ants: ants did not feed on seeds in cones, and so did not compete with cone-foraging birds. However, foraging wood ants did feed on arthropods living in the foliage, thus reducing the amount of food available to birds there.     

Phylogenetic patterns of ant–fungus associations indicate that farming strategies,not only a superior fungal cultivar,explain the ecological success of leafcutter ants

To elucidate fungicultural specializations contributing to ecological dominance of leafcutter ants, we estimate the phylogeny of fungi cultivated by fungus‐growing (attine) ants, including fungal cultivars from (i) the entire leafcutter range from southern South America to southern North America, (ii) all higher‐attine ant lineages (leafcutting genera Atta, Acromyrmex; nonleafcutting genera Trachymyrmex, Sericomyrmex) and (iii) all lower‐attine lineages. Higher‐attine fungi form two clades, Clade‐A fungi (Leucocoprinus gongylophorus, formerly Attamyces) previously thought to be cultivated only by leafcutter ants, and a sister clade, Clade‐B fungi, previously thought to be cultivated only by Trachymyrmex and Sericomyrmex ants. Contradicting this traditional view, we find that (i) leafcutter ants are not specialized to cultivate only Clade‐A fungi because some leafcutter species ranging across South America cultivate Clade‐B fungi; (ii) Trachymyrmex ants are not specialized to cultivate only Clade‐B fungi because some Trachymyrmex species cultivate Clade‐A fungi and other Trachymyrmex species cultivate fungi known so far only from lower‐attine ants; (iii) in some locations, single higher‐attine ant species or closely related cryptic species cultivate both Clade‐A and Clade‐B fungi; and (iv) ant–fungus co‐evolution among higher‐attine mutualisms is therefore less specialized than previously thought. Sympatric leafcutter ants can be ecologically dominant when cultivating either Clade‐A or Clade‐B fungi, sustaining with either cultivar‐type huge nests that command large foraging territories; conversely, sympatric Trachymyrmex ants cultivating either Clade‐A or Clade‐B fungi can be locally abundant without achieving the ecological dominance of leafcutter ants. Ecological dominance of leafcutter ants therefore does not depend primarily on specialized fungiculture of L. gongylophorus (Clade‐A), but must derive from ant–fungus synergisms and unique ant adaptations.     

Specific, non-nutritional association between an ascomycete fungus and Allomerus plant-ants

Ant-fungus associations are well known from attine ants, whose nutrition is based on a symbiosis with basidiomycete fungi. Otherwise, only a few non-nutritional ant-fungus associations have been recorded to date. Here we focus on one of these associations involving Allomerus plant-ants that build galleried structures on their myrmecophytic hosts in order to ambush prey. We show that this association is not opportunistic because the ants select from a monophyletic group of closely related fungal haplotypes of an ascomycete species from the order Chaetothyriales that consistently grows on and has been isolated from the galleries. Both the ants' behaviour and an analysis of the genetic population structure of the ants and the fungus argue for host specificity in this interaction. The ants' behaviour reveals a major investment in manipulating, growing and cleaning the fungus. A molecular analysis of the fungus demonstrates the widespread occurrence of one haplotype and many other haplotypes with a lower occurrence, as well as significant variation in the presence of these fungal haplotypes between areas and ant species. Altogether, these results suggest that such an interaction might represent an as-yet undescribed type of specific association between ants and fungus in which the ants cultivate fungal mycelia to strengthen their hunting galleries.     

Studies on fungi cultivated by ants

A collection of 36 fungi cultivated by leaf-cutting ants has been established at The New York Botanical Garden. These fungi grow on a variety of natural media and on a synthetic medium with mineral salts, dextrose, casein hydrolysate, purine and pyrimidine bases and vitamins. Tests of the fungi for antibacterial activity were all negative againstStaphylococcus aureus andEscherichia coli. Only four isolates of ant fungi, each cultivated by a different species of ant, produced basidiocarps on oatmeal agar. Taxonomic studies indicate that these belong to the same species of fungus (Lepiota sp.). Eighteen isolates produced bromatia characteristic of the form species,Attamyces bromatificus Kreisel, one produced a mycelium with clamp connections, and thirteen produced sterile mycelia without clamped hyphae and without bromatia.     

What determines conditionality in ant–Hemiptera interactions? Hemiptera habitat preference and the role of local ant activity

Abstract.  1. This paper describes spatial variation in density of a mutualist Hemiptera, and attempts to elucidate an understanding of the spatial variation in conditionality of its mutualism with ants.
2. Aggregations of the membracid treehopper, Campylenchia sp., occurred more frequently, and with higher number of individuals, in isolated trees as compared with their occurrence in the same tree species in wooded patches.
3. Using treehopper aggregations as the level of replication, there was a habitat × ant presence interaction associated with colony survival time. However, when the median survival time of all aggregations within each tree was used, only an additive effect of both habitat type and ant presence were apparent. This suggests that treehopper aggregations in some isolated trees experienced more benefit from ant presence than other trees.
4. The ant:treehopper ratio averaged over each tree was correlated with effect of ant presence on survivorship (survival time without ants/survival time with ants). Therefore, conditionality in the ant–membracid mutualism appears related to ant attendance level at the scale of individual trees.
5. Ant attendance at treehopper aggregations in isolated and matorral trees did not differ, and therefore there is no clear mechanism by which ants create the contrast in treehopper densities between the two habitat types.
6. The main force shaping initial treehopper densities in the two habitats is likely plant quality. Results suggest that growing situation (or more specifically plant health) is likely a good predictor of membracid density, while general ant activity within a tree is the best indicator of interaction strength.     

The influence of thermoregulation on behavioural recovery from exercise in a lizard

1. Past work on the thermal preferences of Dipsosaurus dorsalis (Biard & Giard) has indicated that intense, exhaustive exercise causes these lizards to select a body temperature (33·5 °C) which is cooler than their preferred activity temperature of 40°C during the first 1–2 h of exercise recovery.
2. In order to test the hypothesis that the thermal regime selected by exhausted D. dorsalis is beneficial to the process of exercise recovery, lizards were forced to undergo both exhaustive and sprinting exercise at their preferred body temperature of 40°C. The peak speeds attained and the total distances travelled by these animals during these two different exercise protocols were measured and the animals were then forced to undergo a second bout of either sprinting or exhaustive exercise, following a 30–330 min recovery at either 20°C, 40°C or under a variable thermal regime which duplicated that selected by animals following exercise.
3. Animals recovering at a constant 40°C regained their ability to repeat exhaustive activity in less than 85 min, while animals recovering under the other two thermal regimes required between 85 and 100 min of recovery to be able to repeat this activity. Animals recovering at both 40°C and under the variable thermal regime regained their ability to repeat sprint behaviour within 60 min of recovery, while animals recovering at 20°C required more than 100 min of recovery to be able to repeat sprint behaviour.
4. These results formed the basis of the conclusion that the post-exercise behaviour selected by D. dorsalis retards the rate at which the animals recover their ability to repeat exhaustive exercise when compared with recovery at a constant 40°C but does not retard their ability to repeat sprint exercise.     

Selective isolation of dematiaceous fungi from the workers of <Emphasis Type="Italic">Atta laevigata</Emphasis> (Formicidae: Attini)

Leaf-cutting ants (Formicidae: Attini) are considered pests in agriculture for their impact in human crops, as they utilize leaf fragments to raise their fungal mutualist (Agaricales: Lepiotaceae). Basically, the basidiomycetous fungus is cultivated to supply food to adult workers and broads; in return, the ants protect it against natural enemies. However, recent studies have claimed that other microorganisms are associated to ant nests where a wide range of interactions may take place. To investigate the occurrence of dematiaceous fungi on the cuticle of Atta laevigata ants, 30 workers were sampled from an adult nest located in the surroundings of the Center for the Studies of Social Insects, UNESP-Rio Claro, SP, Brazil. The use of selective techniques to avoid high-sporulation fungi has been recommended and was tested in this study. To favor the isolation of the desired fungi, heads and cuticle scrapings of ant bodies were inoculated on Mycosel agar and incubated for 3 weeks at 35°C. Morphological and molecular methods were used to identify the filamentous fungi recovered. From 56 isolates, 19 were hyaline filamentous species, and among the remaining 37, some are mentioned as phyto-associated fungi like Alternaria arborescens, Bipolaris sorokiniana, Bipolaris eleusines, Bipolaris zeae, Curvularia trifolii, and Paraphaeosphaeria michotii. These species are reported from A. laevigata bodies for the first time. None of the isolation trials revealed the presence of the parasite Escovopsis or entomopathogenic fungi. The possible spread of the fungi in nature by the ants is discussed.     

Behavioural and chemical studies of discrimination processes in the leaf-cutting ant Acromyrmex laticeps nigrosetosus (Forel, 1908).

Leaf-cutting ants live in symbiosis with a basidiomycete fungus that is exploited as a source of nutrients for ant larvae. Tests of brood transport revealed that Acromyrmex laticeps nigrosetosus workers did not discriminate a concolonial brood from an alien brood. The same result was observed with tests of fungus transport. Adult workers showed no aggressive behaviour to workers from other alien colonies (non-nestmates). There was no qualitative variation in the chemical profiles of larvae, pupae and adult workers from the different colonies. However, quantitative differences were observed between the different colonies. Hypotheses about the lack of intraspecific aggression in this subspecies of ants are discussed.