Evolution of ant-cultivar specialization and cultivar switching in Apterostigma fungus-growing ants

Almost all of the more than 200 species of fungus-growing ants (Formicidae: Attini) cultivate litter-decomposing fungi in the family Lepiotaceae (Basidiomycota: Agaricales). The single exception to this rule is a subgroup of ant species within the lower attine genus Apterostigma, which cultivate pterulaceous fungi distantly related to the Lepiotaceae. Comparison of cultivar and ant phylogenies suggests that a switch from lepiotaceous to pterulaceous fungiculture occurred only once in the history of the fungus-growing ants. This unique switch occurred after the origin of the genus Apterostigma, such that the basal Apterostigma lineages retained the ancestral attine condition of lepiotaceous fungiculture, and none of the Apterostigma lineages in the monophyletic group of pterulaceous fungiculturists are known to have reverted back to lepiotaceous fungiculture. The origin of pterulaceous fungiculture in attine ants may have involved a unique transition from the ancestral cultivation of litter-decomposing lepiotaceous fungi to the cultivation of wood-decomposing pterulaceous fungi. Phylogenetic analyses further indicate that distantly related Apterostigma ant species sometimes cultivate the same cultivar lineage, indicating evolutionarily frequent, and possibly ongoing, exchanges of fungal cultivars between Apterostigma ant species. The pterulaceous cultivars form two sister clades, and different Apterostigma ant lineages are invariably associated with, and thus specialized on, only one of the two cultivar clades. However, within clades Apterostigma ant species are able to switch between fungi. This pattern of broad specialization by attine ants on defined cultivar clades, coupled with flexible switching between fungi within cultivar clades, is also found in other attine lineages and appears to be a general phenomenon of fungicultural evolution in all fungus-growing ants.     

Convergent coevolution in the domestication of coral mushrooms by fungus-growing ants

Comparisons of phylogenetic patterns between coevolving symbionts can reveal rich details about the evolutionary history of symbioses. The ancient symbiosis between fungus-growing ants, their fungal cultivars, antibiotic-producing bacteria and cultivar-infecting parasites is dominated by a pattern of parallel coevolution, where the symbionts of each functional group are members of monophyletic groups. However, there is one outstanding exception in the fungus-growing ant system, the unidentified cultivar grown only by ants in the Apterostigma pilosum group. We classify this cultivar in the coral-mushroom family Pterulaceae using phylogenetic reconstructions based on broad taxon sampling, including the first mushroom collected from the garden of an ant species in the A. pilosum group. The domestication of the pterulaceous cultivar is independent from the domestication of the gilled mushrooms cultivated by all other fungus-growing ants. Yet it has the same overall assemblage of coevolved ant-cultivar-parasite-bacterium interactions as the other ant-grown fungal cultivars. This indicates a pattern of convergent coevolution in the fungus-growing ant system, where symbionts with both similar and very different evolutionary histories converge to functionally identical interactions.     

Novel mandibular gland volatiles from Apterostigma ants

The fungus-farming ants are a well-studied evolutionary radiation within the subfamily Myrmicinae that associate with a web of symbionts that span kingdoms. Members of the Apterostigma pilosum species group cultivate unique basidiomycete fungi belonging to the coral-mushroom family Pterulaceae, a family of fungi that is distantly related to the Agaricaceae (Leucoagaricus and Leucocoprinus) fungi grown by most fungus-farmers including other members in the genus Apterostigma (A. auriculatum group and A. megacephala). A chemical analysis using gas chromatography–mass spectroscopy of the mandibular gland volatiles of two species – A. dentigerum and A. manni – revealed the presence of an extraordinary diversity of natural products. Many of these compounds are new to Arthropoda, such as a homologous series of 3-methyl-2-alkanones, 2-methyl-2-alkenals, and 1-phenyl-2-propanone in A. dentigerum and 1-phenyl-2-propanol in A. manni. These results identify a remarkable divergence of compounds across the fungus-growing ants and other members in Myrmicinae. Functions of these natural products are proposed and discussed.     

Frontier mutualism: coevolutionary patterns at the northern range limit of the leaf-cutter ant-fungus symbiosis

Tropical leaf-cutter ants cultivate the fungus Attamyces bromatificus in a many-to-one, diffuse coevolutionary relationship where ant and fungal partners re-associate frequently over time. To evaluate whether ant-Attamyces coevolution is more specific (tighter) in peripheral populations, we characterized the host-specificities of Attamyces genotypes at their northern, subtropical range limits (southern USA, Mexico and Cuba). Population-genetic patterns of northern Attamyces reveal features that have so far not been observed in the diffusely coevolving, tropical ant-Attamyces associations. These unique features include (i) cases of one-to-one ant-Attamyces specialization that tighten coevolution at the northern frontier; (ii) distributions of genetically identical Attamyces clones over large areas (up to 81 000 km(2), approx. the area of Ireland, Austria or Panama); (iii) admixture rates between Attamyces lineages that appear lower in northern than in tropical populations; and (iv) long-distance gene flow of Attamyces across a dispersal barrier for leaf-cutter ants (ocean between mainland North America and Cuba). The latter suggests that Attamyces fungi may occasionally disperse independently of the ants, contrary to the traditional assumption that Attamyces fungi depend entirely on leaf-cutter queens for dispersal. Peripheral populations in Argentina or at mid-elevation sites in the Andes may reveal additional regional variants in ant-Attamyces coevolution. Studies of such populations are most likely to inform models of coextinctions of obligate mutualistic partners that are doubly stressed by habitat marginality and by environmental change.     

Genetic diversity of termite-egg mimicking fungi “termite balls” within the nests of termites

Antagonistic or mutualistic interactions between insects and fungi are well-known, and the mutualistic interactions of fungus-growing ants, fungus-growing termites, and fungus-gardening beetles with their respective fungal mutualists are model examples of coevolution. However, our understanding of coevolutionary interactions between insects and fungi has been based on a few model systems. Fungal mimicry of termite eggs is one of the most striking evolutionary consequences of insect–fungus associations. This novel termite–fungus interaction is a good model system to compare with the relatively well-studied systems of fungus-growing ants and termites because termite egg-mimicking fungi are protected in the nests of social insects, as are fungi cultivated by fungus-growing ants and termites. Recently, among systems of fungus-growing ants and termites, much attention has been focused on common factors including monoculture system for the ultimate evolutionary stability of mutualism. We examined the genetic diversity of termite egg-mimicking fungi within host termite nests. RFLP analysis demonstrated that termite nests were often infected by multiple strains of termite egg-mimicking fungi, in contrast to single-strain monocultures in fungus combs of fungus-growing ants and termites. Additionally, phylogenetic analyses indicated the existence of a free-living stage of the termite egg-mimicking fungus as well as frequent long-distance gene flow by spores and subsequent horizontal transmission. Comparisons of these results with previous studies of fungus-growing ants and termites suggest that the level of genetic diversity of fungal symbionts within social insect nests may be important in shaping the outcome of the coevolutionary interaction, despite the fact that the mechanism for achieving genetic diversity varies with the evolutionary histories of the component species.     

Phylogenetic analysis of mutualistic filamentous bacteria associated with fungus-growing ants

The attine ant-microbe system is a quadripartite symbiosis, involving a complex set of mutualistic and parasitic associations. The symbiosis includes the fungus-growing ants (tribe Attini), the basidiomycetous fungi the ants cultivate for food, specialized microfungal parasites (in the genus Escovopsis) of the cultivar, and ant-associated mu tualistic filamentous bacteria that secrete antibiotics specifically targeted to suppress the growth of Escovopsis. In this study, we conduct the first phylogenetic analysis of the filamentous mutualistic bacteria (actinomycetes) associated with fungus-growing ants. The filamentous bacteria present on 3 genera of fungus-growing ants (Acromyrmex, Trachy myrmex, and Apterostigma) were isolated from 126 colonies. The isolated actinomycetes were grouped into 3 distinct morphological types. Each morphological type was specific to the ant genus from which it was isolated, suggesting some degree of host specificity. The phylogenetic position of the 3 morphotypes was estimated using 16S rDNA for representative strains. The 8 isolates of actinomycetes sequenced are in the family Pseudonocardiaceae (Actino mycetales) and belong to the genus Pseudonocardia. Transmission electron microscopy examination of the actino mycete associated with the cuticle of Acromyrmex sp. revealed bacterial cells with an outer electron-dense membrane, consistent with actinomycetes in the genus Pseudonocardia. Ant-associated Pseudonocardia isolates did not form a monophyletic group, suggesting multiple acquisitions of actinomycetes by fungus-growing ants over their evolutionary history.     

Coevolution between attine ants and actinomycete bacteria: a reevaluation

We reassess the coevolution between actinomycete bacteria and fungus-gardening (attine) ants. Actinomycete bacteria are of special interest because they are metabolic mutualists of diverse organisms (e.g., in nitrogen-fixation or antibiotic production) and because Pseudonocardia actinomycetes are thought to serve disease-suppressing functions in attine gardens. Phylogenetic information from culture-dependent and culture-independent microbial surveys reveals (1) close affinities between free-living and ant-associated Pseudonocardia, and (2) essentially no topological correspondence between ant and Pseudonocardia phylogenies, indicating frequent bacterial acquisition from environmental sources. Identity of ant-associated Pseudonocardia and isolates from soil and plants implicates these environments as sources from which attine ants acquire Pseudonocardia. Close relatives of Atta leafcutter ants have abundant Pseudonocardia, but Pseudonocardia in Atta is rare and appears at the level of environmental contamination. In contrast, actinomycete bacteria in the genera Mycobacterium and Microbacterium can be readily isolated from gardens and starter-cultures of Atta. The accumulated phylogenetic evidence is inconsistent with prevailing views of specific coevolution between Pseudonocardia, attine ants, and garden diseases. Because of frequent acquisition, current models of Pseudonocardia-disease coevolution now need to be revised. The effectiveness of Pseudonocardia antibiotics may not derive from advantages in the coevolutionary arms race with specialized garden diseases, as currently believed, but from frequent recruitment of effective microbes from environmental sources. Indeed, the exposed integumental structures that support actinomycete growth on attine ants argue for a morphological design facilitating bacterial recruitment. We review the accumulated evidence that attine ants have undergone modifications in association with actinomycete bacteria, but we find insufficient support for the reverse, modifications of the bacteria resulting from the interaction with attine ants. The defining feature of coevolution--reciprocal modification--therefore remains to be established for the attine ant-actinomycete mutualism.     

Rectal enzymes of attine ants. α-Amylase and chitinase

The faecal material of seven species of attine ants from the genera Cyphomyrmex, Apterostigma, Myrmicocrypta, Sericomyrmex, and Atta has been shown to contain α-amylase and chitinase, but only a trace of uricase. Chitinase probably serves a beneficial rôle in the fungus-culturing activities of the primitive forms by contributing to the degradation of chitinous substrates, such as insect cuticle, and by lysing potentially competitive chitinous fungi. Biochemical factors significant in the evolution of the fungus-growing ants are discussed.     

Instability of novel ant-fungal associations constrains horizontal exchange of fungal symbionts

One of the more fascinating features of fungus-gardening ants (Attini: Formicidae) is their fidelity to their lineage-specific fungal symbionts. Among the derived higher-attine ants (leafcutter ants and close relatives), it is thought that most leaf-cutting ants grow Attamyces fungus whereas most Trachymyrmex ants grow ‘Trachymyces’ fungus, but there exist exceptions to this clade-to-clade correspondence between ants and fungi. The exceptions are inconsistent with strict one-to-one coevolution, which suggests that ants sometimes are able to switch to novel fungi. Such switches appear to be largely constrained and ants are generally faithful to their species-specific fungi. Prior experiments demonstrated no clear fitness consequences of growing novel fungi over the short-term when the ant Trachymyrmex septentrionalis was symbiont-switched by forcing it to grow Attamyces leaf-cutter fungus. We hypothesized that long-term ant-fungal fidelity is constrained either by physiological differences among fungal species or by garden diseases that symbiont-switched ants cannot control. Repeat experiments in a different location show that T. septentrionalis colonies switched to grow Attamyces exhibit sudden declines in garden biomass and consequent fitness reductions due to garden destruction by pathogens, whereas control colonies (Trachymyrmex ants cultivating Trachymyces fungus) do not show parallel garden declines. These patterns are mirrored in symbiont-switch experiments conducted on colonies in Trachymyrmex turrifex. Disease microbes selecting on ant-cultivar combinations therefore can constrain switches to novel cultivars and maintain combinations that are more resistant to disease.     

On the run: free-living mushroom corals avoiding interaction with sponges

Individuals of the free-living mushroom coral Heliofungia fralinae moved away when placed in contact with fragments of the toxic haplosclerid sponge Callyspongia (Euplacella) biru. This reaction was not evoked by three other sponge species. The experiment demonstrated that mobility of mushroom corals helps them to flee from organisms that secrete secondary metabolites in competition for space.     

Extensive exchange of fungal cultivars between sympatric species of fungus-growing ants

Fungal cultivars of fungus-growing ants (Attini, Formicidae) are carried by dispersing queens from parent to offspring nest. This vertical cultivar transmission between generations is thought to result in long-term ant-fungus coevolution and selection for beneficial cultivar traits that maximize harvests and thus colony productivity. In contrast to this traditional view of vertical cultivar transmission, frequent horizontal cultivar transmission between ant species is implicated by a phylogenetic analysis of 72 cultivars propagated by two fungus-growing ant species coexisting sympatrically in central Panama. The two ant species are specialized on the same group of closely related cultivars, but in six of 12 cultivar clades identifiable within this group, cultivars from both ant species were united in the same clade. Five of these 'mixed' clades were supported by bootstrap values of about 90% or higher. In one instance, colonies from the two ant species cultivated the same, genetically identical, cultivar clone. These phylogenetic patterns indicate that: (i) cultivar exchanges between the two ant species occur routinely throughout ecological time; and that (ii) coevolutionary processes between ants and their fungi are more diffuse than previously assumed. Because the two ant species are specialized on a narrow group of closely related cultivars that they regularly exchange among each other, but not with other sympatric ant species, cultivar exchanges are constrained, most likely, by ant preferences for their own cultivar group or by stringent selection against transitions of ant lineages to distantly related cultivars.     

Specificity in the symbiotic association between fungus-growing ants and protective Pseudonocardia bacteria

Fungus-growing ants (tribe Attini) engage in a mutualism with a fungus that serves as the ants' primary food source, but successful fungus cultivation is threatened by microfungal parasites (genus Escovopsis). Actinobacteria (genus Pseudonocardia) associate with most of the phylogenetic diversity of fungus-growing ants; are typically maintained on the cuticle of workers; and infection experiments, bioassay challenges and chemical analyses support a role of Pseudonocardia in defence against Escovopsis through antibiotic production. Here we generate a two-gene phylogeny for Pseudonocardia associated with 124 fungus-growing ant colonies, evaluate patterns of ant-Pseudonocardia specificity and test Pseudonocardia antibiotic activity towards Escovopsis. We show that Pseudonocardia associated with fungus-growing ants are not monophyletic: the ants have acquired free-living strains over the evolutionary history of the association. Nevertheless, our analysis reveals a significant pattern of specificity between clades of Pseudonocardia and groups of related fungus-growing ants. Furthermore, antibiotic assays suggest that despite Escovopsis being generally susceptible to inhibition by diverse Actinobacteria, the ant-derived Pseudonocardia inhibit Escovopsis more strongly than they inhibit other fungi, and are better at inhibiting this pathogen than most environmental Pseudonocardia strains tested. Our findings support a model that many fungus-growing ants maintain specialized Pseudonocardia symbionts that help with garden defence.     

蚂蚁对澜沧舞花姜种子散布及种苗空间分布格局的影响

 在野外系统观测了澜沧舞花姜（Globba lancangensis）18个果实共216粒种子的散布过程。共有10种蚂蚁参与了澜沧舞花姜的种子散布,距离为0.01～3.35 m,平均距离(0.47±0.03) m（平均值±SE,n=216）。其中最重要的３种蚂蚁是横纹齿猛蚁（Odontoponera transversa）、大头蚁(Pheidole sp.)和黄足厚结猛蚁(Pachycondyla luteipes),其出现频率分别为61%、50%和28%,散布的平均距离分别为(0.60±0.09) m、(0.20±0.01) m和(0.32±0.05) m。从总体上看,蚂蚁促进了种子的分散,降低了种子的聚集程度。横纹齿猛蚁对于种子上的油质体最为敏感,对人工去除了油质体的种子不搬运,对种子散布距离较远,暗示了其与澜沧舞花姜之间可能存在更紧密的互惠关系。野外样方调查结果表明,在3种舞花姜属植物中,以种子繁殖为主的澜沧舞花姜种苗之间的平均最近距离为(36.8±1.45) cm（平均值±SE,n=74）,显著大于以珠芽繁殖为主的毛舞花姜（Globba barthiri）的(29.8±2.70) m（n=34）（t73,33=2.11,p=0.037）和异果舞花姜（Globba racemosa）的(28.7±3.16) cm（n=32）（t73,31=2.33,p=0.022）;澜沧舞花姜的种苗聚集程度（Z=-1.70±0.19）显著小于毛舞花姜（Z=-2.58±0.37,t73,33=2.36,p=0.020）和异果舞花姜（Z=-3.28±0.53）（t73,31=3.54,p=0.001）。这说明相对于毛舞花姜和异果舞花姜,蚂蚁对种子的散布作用显著增加了澜沧舞花姜种苗间的平均最近距离,降低了居群的聚集度。     

Low host-pathogen specificity in the leaf-cutting ant-microbe symbiosis

Host-parasite associations are shaped by coevolutionary dynamics. One example is the complex fungus-growing ant-microbe symbiosis, which includes ancient host-parasite coevolution. Fungus-growing ants and the fungi they cultivate for food have an antagonistic symbiosis with Escovopsis, a specialized microfungus that infects the ants' fungus gardens. The evolutionary histories of the ant, cultivar and Escovopsis are highly congruent at the deepest phylogenetic levels, with specific parasite lineages exclusively associating with corresponding groups of ants and cultivar. Here, we examine host-parasite specificity at finer phylogenetic levels, within the most derived clade of fungus-growing ants, the leaf-cutters (Atta spp. and Acromyrmex spp.). Our molecular phylogeny of Escovopsis isolates from the leaf-cutter ant-microbe symbiosis confirms specificity at the broad phylogenetic level, but reveals frequent host-switching events between species and genera of leaf-cutter ants. Escovopsis strains isolated from Acromyrmex and Atta gardens occur together in the same clades, and very closely related strains can even infect the gardens of both ant genera. Experimental evidence supports low host-parasite specificity, with phylogenetically diverse strains of Escovopsis being capable of overgrowing all leaf-cutter cultivars examined. Thus, our findings indicate that this host-pathogen association is shaped by the farming ants having to protect their cultivated fungus from phylogenetically diverse Escovopsis garden pathogens.     

Dietary shift in corallivorous Drupella snails following a major bleaching event at Koh Tao, Gulf of Thailand

The island Koh Tao in the western Gulf of Thailand suffered severe coral bleaching in 2010. Its mushroom coral fauna of 20 species was surveyed during the bleaching in 2010 and after the bleaching in 2011. Multi-species assemblages of free-living mushroom corals occurred around the island, two of which were invaded by corallivorous Drupella snails after the bleaching. Previously these gastropods were known to mainly consume branching corals and hardly any mushroom corals. The snails were found preying on four fungiid species, three of which were susceptible to bleaching. The dietary shift became apparent after populations of preferred prey species (Acroporidae and Pocilloporidae) had died during the bleaching event. It seems that bleaching mortality reduced the availability of preferred prey, causing the corallivores to switch to less preferred species that occur in dense aggregations.     

Evolutionary transition from single to multiple mating in fungus-growing ants

Queens of leafcutter ants exhibit the highest known levels of multiple mating (up to 10 mates per queen) among ants. Multiple mating may have been selected to increase genetic diversity among nestmate workers, which is hypothesized to be critical in social systems with large, long-lived colonies under severe pressure of pathogens. Advanced fungus-growing (leafcutter) ants have large numbers (104-106 workers) and long-lived colonies, whereas basal genera in the attine tribe have small (< 200 workers) colonies with probably substantially shorter lifespans. Basal attines are therefore expected to have lower queen mating frequencies, similar to those found in most other ants. We tested this prediction by analysing queen mating frequency and colony kin structure in three basal attine species: Myrmicocrypta ednaella, Apterostigma collare and Cyphomyrmex longiscapus. Microsatellite marker analyses revealed that queens in all three species were single mated, and that worker-to-worker relatedness in these basal attine species is very close to 0.75, the value expected under exclusively single mating. Fungus growing per se has therefore not selected for multiple queen mating. Instead, the advanced and highly productive social structure of the higher attine ants, which is fully dependent on the rearing of an ancient clonal fungus, may have necessitated high genetic diversity among nestmate workers. This is not the case in the lower attines, which rear fungi that were more recently derived from free-living fungal populations.     

Identifying the transition between single and multiple mating of queens in fungus-growing ants

Obligate mating of females (queens) with multiple males has evolved only rarely in social Hymenoptera (ants, social bees, social wasps) and for reasons that are fundamentally different from those underlying multiple mating in other animals. The monophyletic tribe of ('attine') fungus-growing ants is known to include evolutionarily derived genera with obligate multiple mating (the Acromyrmex and Atta leafcutter ants) as well as phylogenetically basal genera with exclusively single mating (e.g. Apterostigma, Cyphomyrmex, Myrmicocrypta). All attine genera share the unique characteristic of obligate dependence on symbiotic fungus gardens for food, but the sophistication of this symbiosis differs considerably across genera. The lower attine genera generally have small, short-lived colonies and relatively non-specialized fungal symbionts (capable of living independently of their ant hosts), whereas the four evolutionarily derived higher attine genera have highly specialized, long-term clonal symbionts. In this paper, we investigate whether the transition from single to multiple mating occurred relatively recently in the evolution of the attine ants, in conjunction with the novel herbivorous 'leafcutter' niche acquired by the common ancestor of Acromyrmex and Atta, or earlier, at the transition to rearing specialized long-term clonal fungi in the common ancestor of the larger group of higher attines that also includes the genera Trachymyrmex and Sericomyrmex. We use DNA microsatellite analysis to provide unambiguous evidence for a single, late and abrupt evolutionary transition from exclusively single to obligatory multiple mating. This transition is historically correlated with other evolutionary innovations, including the extensive use of fresh vegetation as substrate for the fungus garden, a massive increase in mature colony size and morphological differentiation of the worker caste.     

A phylogenetic analysis of the fungus-growing ants (Hymenoptera: Formicidae: Attini) based on morphological characters of the larvae

A phylogenetic hypothesis of the fungus-growing ants (subfamily Myrmicinae, tribe Attini) is proposed, based on a cladistic analysis utilizing forty-four morphological characters (109 states) of the prepupal worker larva. The fifty-one attine species analysed include representatives of eleven of the twelve currently recognized attine genera, excluding only the monotypic workerless parasite Pseudoatta ; the non-attines include two outgroups (species of the basal myrmicine genera Myrmica and Pogonomyrmex ), two myrmicine species presumed to be distantly related to the attines, and twelve species representing taxa that have been proposed by prior workers as possible sister groups of the Attini. There is strong character support for the monophyly of the Attini and for a sister-group relationship of the Attini and the Neotropical Blepharidatta brasiliensis. The Attini are divided into two distinct lineages, an 'apterostigmoid' clade (containing Apterostigma and Mycocepurus) and an 'attoid' clade (containing all other attine genera except Myrmicocrypta). The attine genus Myrmicocrypta appears to be paraphyletic with respect to these two groups; the species M.buenzlii in particular retains many attine plesiomorphies.
These results indicate that the fungus-growing behaviour had a single evolutionary origin in the ants. They also indicate that mycelium cultivation is plesiomorphic and that yeast cultivation is derived within the Attini, overturning the long-standing assumption that the yeast-growing Cyphomyrmex species are the most primitive Attini. Behavioural and ecological investigations into the origin and evolution of the fungus-growing behaviour might more profitably focus on species in the attine genus Myrmicocrypta , as well as those in the closely related non-attine genera Blepharidatta and Wasmannia.     

Disease management in two sympatric Apterostigma fungus‐growing ants for controlling the parasitic fungus Escovopsis

Antagonistic interactions between host and parasites are often embedded in networks of interacting species, in which hosts may be attacked by competing parasites species, and parasites may infect more than one host species. To better understand the evolution of host defenses and parasite counterdefenses in the context of a multihost, multiparasite system, we studied two sympatric species, of congeneric fungus‐growing ants (Attini) species and their symbiotic fungal cultivars, which are attacked by multiple morphotypes of parasitic fungi in the genus, Escovopsis. To assess whether closely related ant species and their cultured fungi are evolving defenses against the same or different parasitic strains, we characterized Escovopsis that were isolated from colonies of sympatric Apterostigma dentigerum and A. pilosum. We assessed in vitro and in vivo interactions of these parasites with their hosts. While the ant cultivars are parasitized by similar Escovopsis spp., the frequency of infection by these pathogens differs between the two ant species. The ability of the host fungi to suppress Escovopsis growth, as well as ant defensive responses toward the parasites, differs depending on the parasite strain and on the host ant species.