A geographical mosaic of coevolution in a slave-making host-parasite system.

Three different isolated populations of the slave-making ant Rossomyrmex minuchae, sympatric with its obligate host Proformica longiseta, are known from the high mountains of southern Spain. To test the prediction that the slave-maker and its host represent a coevolutionary geographical mosaic, we studied the variation in the cuticular hydrocarbons (CHCs) as the trait most likely to show the selection mosaic, plus trait remixing by the gene flow in the populations of each species by means of microsatellites. We found within populations, host and parasite had more similar CHC profiles than between the populations or between parasites and allopatric hosts. The differences between the CHC profiles of the host and parasite, which may be responsible for the level of tolerance towards the parasite, varied between the populations suggesting the existence of a selection mosaic of coevolution. Furthermore, P. longiseta showed higher gene flow than R. minuchae, which would allow local variation in the coevolution of the host and parasite while allowing some trait remixing.

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.

Behavioural and chemical studies of discrimination processes in the leaf-cutting ant Acromyrmex laticeps nigrosetosus (Forel, 1908) / Estudos químico e comportamental dos processos discriminatórios na formiga cortadeira 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.

As formigas cortadeiras vivem em simbiose com um fungo basidiomiceto que é utilizado como fonte de nutriente para suas larvas. Testes de transporte de prole revelaram que as operárias de Acromyrmex laticeps nigrosetosus não discriminaram a prole concolonial de prole estranha. O mesmo resultado foi verificado com testes de transporte do fungo. As operárias adultas não exibiram comportamento agressivo frente a operárias de outras colônias (não companheiras de ninho). Não houve variação qualitativa nos perfis químicos de larvas, pupas e operárias adultas de diferentes colônias. No entanto, diferenças quantitativas foram observadas entre as diferentes colônias. Hipóteses sobre a ausência de agressão intra-específica nesta subespécie de formiga são discutidas.

Phylogeny of ants (Formicidae) based on morphology and DNA sequence data.

In order to reconstruct ants' phylogeny, we analysed DNA sequences for two nuclear genes, abdominal-A and Ultrabithorax, from 49 species of ants and two outgroups. As these genes control the development of the first segments of the abdomen in insects, which are very variable in ants (petiole, postpetiole, and gaster constriction), we hypothesized that the morphological variations between the subfamilies may be correlated with mutations of some abd-A or Ubx regions. Contrarily to our hypothesis, these sequences are highly conserved. The differences observed concern mainly third codon positions and present some saturation. Phylogenetic reconstructions were carried out using the genetic raw sequence data and by combining them with a set of morphological data (Total Evidence). Relations among subfamilies of ants remains poorly resolved with molecular data only, but adding these data to morphological characters confirms and reinforce the topology of : a Poneroid complex [Ponerinae, Cerapachyinae, Leptanillinae and army ants], a Formicoid complex [Dolichoderinae, Formicinae] and a Myrmecoid complex [Myrmicinae, Myrmeciinae, Pseudomyrmecinae, Nothomyrmeciinae]. Our molecular results allow resolution near the branch tips and three subfamilies (Dolichoderinae, Formicinae and Pseudomyrmecinae) always appear as monophyletic. The Formicinae and the Dolichoderinae have close relationships. The Camponotini appear as a strong clade inside the Formicinae. The Ponerinae are separated in two parts: the Ectatommini and all other tribes. The Cerapachyinae, Dorylinae, and Ecitoninae belong to the same clade, the Cerapachyinae being confirmed in their subfamily status. The Myrmicinae appears to be very heterogeneous, with the Attini forming a very stable and well-separated group.

Blending in with the crowd: social parasites integrate into their host colonies using a flexible chemical signature.

Social parasites are able to exploit their host's communication code and achieve social integration. For colony foundation, a newly mated slave-making ant queen must usurp a host colony. The parasite's brood is cared for by the hosts and newly eclosed slave-making workers integrate to form a mixed ant colony. To elucidate the social integration strategy of the slave-making workers, Polyergus rufescens, behavioural and chemical analyses were carried out. Cocoons of P. rufescens were introduced into subcolonies of four potential host species: Formica subgenus Serviformica (Formica cunicularia and F. rufibarbis, usual host species; F. gagates, rare host; F. selysi, non-natural host). Slave-making broods were cared for and newly emerged workers showed several social interactions with adult Formica. We recorded the occurrence of abdominal trophallaxis, in which P. rufescens, the parasite, was the donor. Social integration of P. rufescens workers into host colonies appears to rely on the ability of the parasite to modify its cuticular hydrocarbon profile to match that of the rearing species. To study the specific P. rufescens chemical profile, newly emerged callows were reared in isolation from the mother colony (without any contact with adult ants). The isolated P. rufescens workers exhibited a chemical profile closely matching that of the primary host species, indicating the occurrence of local host adaptation in the slave-maker population. However, the high flexibility in the ontogeny of the parasite's chemical signature could allow for host switching.

The role of the symbiotic fungus in the digestive metabolism of two species of fungus-growing ants.

Leaf-cutting ants live in an obligatory symbiosis with a fungus which they grow on fresh leaves harvested by workers. This study attempts to clarify the respective role of ants and fungus in the degradation of plant material, in order to highlight the evolutionary basis of this mutualistic association. The symbiotic system of two ant species, Acromyrmex subterraneus subterraneus and Acromyrmex crassispinus, was investigated. To identify the digestive carbohydrases, a total of 19 specific and synthetic plant material substrates were tested on workers from different castes (major and minor), larvae and fungus. Extracts of A. subterraneus and A. crassispinus workers showed high enzymatic activity particularly on starch, maltose, sucrose and alpha-1,4 glucoside. Larvae degraded starch, sucrose, maltose but also laminarin, and all the detected activities were higher than those found for workers. The symbiotic fungus of A. subterraneus was mostly active on laminarin, xylan and cellulose, while the symbiotic fungus of A. crassispinus was mostly active on laminarin, starch, maltose and sucrose. The enzymatic activities of ants and fungus belonging to the same symbiotic system tended not to overlap, suggesting that the association is highly evolved and of an ancient origin.

Lack of intraspecific aggression in the ant Tetramorium bicarinatum: a chemical hypothesis.

Tetramorium bicarinatum (Myrmicinae) is an ant species frequently found in tropical and subtropical areas, particularly in Africa, Southeast Asia (Japan), and South America (Brazil). The species is polygynous, reproduces by budding, and has sterile workers. Since the nests are widely distributed in a given area, the problem arises of territorial defense against conspecifics. Because not all ants defend territories, we assessed the defensive behavior of T. bicarinatum workers through intraspecific and interspecific aggressiveness tests. A detailed behavioral study of the interactions between workers from several different colonies of T. bicarinatum (originating from Japan and Brazil) showed that workers do not discriminate against conspecific nonnestmate individuals, but they are highly aggressive towards allospecifics (Myrmica rubra, Myrmicinae). The results suggest that each colony from this ant species possesses a similar colonial odor. Chemical analyses of the cuticular hydrocarbons of these species were made with gas chromatography coupled to mass spectrometry. Results showed that the different colonies of T. bicarinatum possess a common chemical profile mainly composed of straight-chain alkanes and alkenes, while M. rubra possess more methyl-branched alkanes. We suggest that methyl alkane cues play a determining role in colonial recognition and that these results could explain the underlaying basis of the lack of intraspecific aggressiveness in T. bicarinatum.

Chemical ecology and social parasitism in ants.

The chemical strategies by which parasites manage to break into the social fortresses of ants offer a fascinating theme in chemical ecology. Semiochemicals used for interindividual nestmate recognition are also involved in the mechanisms of tolerance and association between the species, and social parasites exploit these mechanisms. The obligate parasites are odorless ("chemical insignificance") at the time of usurpation, like all other callow ants, and this "invisibility" enables their entry into the host colony. By chemical mimicry (sensu lato), they later integrate the gestalt odor of this colony ("chemical integration"). We hypothesize that host and parasite are likely to be related chemically, thereby facilitating the necessary mimicry to permit bypassing the colony odor barrier. We also review the plethora of chemical weapons used by social parasites (propaganda, appeasement, and/or repellent substances), particularly during the usurpation period, when the young mated parasite queen synthesizes these chemicals before usurpation and ceases such biosynthesis afterwards. We discuss evolutionary trends that may have led to social parasitism, focusing on the question of whether slave-making ants and their host species are expected to engage in a coevolutionary arms race.

Induced mimicry of colony odors in ants.

The cuticular hydrocarbons ofFormica selysi (Formicinae) andMonica rubida (Myrmicinae) reared in single species and in mixed species colonies were determined using gas chromatography (GC) and GC-mass spectrometry. In colonies containing both species, each species modified its species-specific recognition odor. This odor is composed, at least partly, of cuticular hydrocarbons. The cuticular hydrocarbons ofM. rubida consist only of saturated alkanes (n-alkanes and branched alkanes). InF. selysi the mixture also contains unsaturated compounds (monoenes and dienes). In hetero-specific colonies, a new chemical signature developed. This signature resulted from qualitative and quantitative changes in the spectrum of hydrocarbons produced by each species and permitted the two species to inhabit the same nest without displaying interspecific aggression. The readjustment seemed to be more an active synthesis or an active transfer than simply a passive transfer from one species to the other. This may imply that the ants synthesized some components of the hydrocarbon signature of the other species. These synthesizing processes may be activated under particular social environmental conditions.

The organization of artificial heterospecific ant colonies. The case of the Manica rubida/Formica selysi association: mixed colony or parallel colonies?

We studied the spatial and social organization of homospecific and heterospecific colonies of two ant species Manica rubida (Myrmicanae) and Formica selysi (Formicinae) reared without a queen in artificial nests. We used a method based on individual labelling and automated photographic recording of data. The two species, which are forced to live together in the same artificial mixed colony, show the same spatial organization as they do in control homospecific colonies. The Manica rubida were mostly clustered in one chamber while Formica selysi workers were spread throughout the nest. Formica selysi individual spatial profiles are very heterogeneous while in the other species they are very homogeneous. By using taxonomical techniques on behavioural data we show that the two species within the mixed colony are clearly distinguishable. An artificial association of these two species may therefore be considered to be a juxtaposition of two colonies functioning in parallel, rather than a "mixed colony".

Phylogenesis / biotope interactions among formicidae.

The development of interspecific recognition and the expression of behavioral plasticity as a function of phyletic distance and the natural environment in which the ants live were investigated. An experimental model involving artificial mixed societies was used in this study. These mixed societies were produced by combining ants from two out of the 11 investigated species. The possibilities thus ranged from situations in which phylogenetically similar ants that live in the same biotype were combined, to those in which the species differ significantly in terms both of phyletic distance and of biotope. The interspecific groups were established in a neutral environment with adult ants less than 24 hours old. The results concerned agonistic and non-agonistic behaviors during the first 15 days. Both the frequency and types of interaction were shown to vary according to the type of group. Two species that share a common habitat are less aggressive and display more non-agonistic behavior than when the species come from different habitats. On the other hand, although aggressive behaviors increase with greater phyletic distance between the species, non-agonistic interactions remain constant. As a general rule, interactions between species from differing environments seem to be determined by the phyletic distance, whereas those between species from the same biotope are controlled by interspecific competition. There is a gradual change in these interactions dependent on the duration of the mixed association.