Contribution to the conservation biology of the wood ant Formica lugubris (Hymenoptera, Formicidae) in Switzerland

Summary Among ants, wood ants are probably the most fascinating and studied species in temperate European forests. Unfortunately, due to several threats they are nowadays registered in red lists. Recent studies made in the Swiss Jura Mountains ended up in the description of a new sympatric sibling species of Formica lugubris (i.e. Formica paralugubris Seifert 1996). Because of this confusion the biology of F. lugubris is incomplete. Due to the extreme difficulties to distinguish morphologically F. lugubris from F. paralugubris we studied their cuticular hydrocarbons profiles. Irrespective of their geographic origin, we observed quantitative discrimination between species within each caste (workers, males and gynes =young alate female). Moreover, using a behavioural taxonomic approach (i.e. the pupa-carrying test) we showed that ants preferred conspecific worker pupae to those of the sibling species. These first results allowed us to consider the two species as two separate taxonomic units. To understand their coexistence, habitat distribution models were fitted with GIS predictors and factors known to influence wood ant distribution. In the Jura Mountains, although the two species share very similar habitats, they are spatially segregated. F. lugubris occurs more frequently at woodland borders than in forest interiors. We demonstrated with genetic and field data that Formica lugubris displays two different social forms in close proximity in alpine zone (e.g. unmanaged forests of the Swiss National Park). We discovered populations mostly monogynous to weakly polygynous (i.e. one to a few egg laying queens per colony) and monodomous (i.e. one nest per colony), and polygynous/polydomous populations (new nests being founded by colony budding). It is generally admitted that monogyne species disperse well in order to find suitable habitat to found new colonies whereas polygyne species have restricted dispersal and local mating within the nest. In order to compare reproductive strategies of F. lugubris and F. paralugubris (i.e. matings and dealation process) we conducted experiments with sexuals. F, lugubris gynes from monogynous/monodomous populations do not show a local strategy like the obligately polygynous F. paralugubris (i.e. early dealation even without mating, insemination without flight activity and low fat reserve). They always keep their wings, do not mate when not able to fly and have high amount of fat content revealing high survival capacities. On the other side, F, lugubris gynes from polygynous/polydomous populations have lower lipid reserves and displayed a reproductive behaviour close to the F. para lugubris one. After dispersal, wood ant gynes can either start new societies by temporary social parasitism of another species (i.e. subgenus Serviformica) or be adopted intraspecifically in an existing nest. In F. lugubris, we demonstrated that gynes from monogynous/monodomous colonies showed a high success for temporary social parasitism compare to the lower success of gynes from polygynous/polydomous colonies. However, physiological analyses suggested that only gynes from monogynous/ monodomous populations can efficiently disperse and found new nest by temporary social parasitism. Intraspecifically, gynes were accepted to a high degree in polygynous nest and in monogynous nests as long as these nests contained sexuals. In conclusion, Formica lugubris displays a social and dispersal polymorphism (mixed mating and founding system) representing a behavioural plasticity in relation to environmental and ecological conditions. Therefore, conservation measures directed toward this species should try to maintain a maximum of diversity at the habitat level. Résumé Les fourmis des bois sont probablement parmi les espèces de fourmis les plus fascinantes et les plus étudiées des forêts tempérées Européennes. Actuellement, du fait de différentes menaces, elles figurent malheureusement sur listes rouges. Plusieurs études menées au sein du Jura Suisse ont abouti à la description d'une nouvelle espèce jumelle et sympatrique de Formica lugubris (F. para- lugubris Seifert 1996). A cause de cette confusion la biologie de F lugubris est lacunaire. La distinction morphologique de F. lugubris et de F. para lugubris est si difficile que nous avons étudié leurs hydrocarbures cuticulaires. Indépendamment de l'origine géographique, nous avons observé une discrimination quantitative entre les espèces au sein de chaque caste (ouvrières, mâles et jeunes femelles ailées). De plus, à l'aide d'une approche taxonomique comportementale (le test de transport de cocons) nous avons montré que les fourmis préfèrent des cocons d'ouvrières conspécifiques à ceux de l'espèce jumelle. Ces premiers résultats nous permettent de considérer ces deux espèces comme deux unités taxonomiques distinctes et valables. Afin de comprendre leur coexistence, des modèles mathématiques ont été développés avec des données SIG et des facteurs écologiques influençant la répartition des fournis des bois. Dans le Jura, même si elles partagent des habitats fortement similaires, les deux espèces n'occupent pas les mêmes secteurs. F. lugubris est plus fréquente en lisière forestière plutôt qu'en pleine forêt. Nous avons démontré grâce à des données génétiques et de terrain que F. lugubris présente deux formes sociales au sein de la zone alpine (forêts protégées du Parc National Suisse). D'autre part, nous avons découvert des populations monogynes à faiblement polygynes (une à quelques reines pondeuses par colonie) et monodomes (colonies composées d'une seule fourmilière), et des populations polygynes/polydomes (les nouveaux nids étant produit par bourgeonnement). Généralement, les espèces monogynes dispersent sur de grandes distances et peuvent coloniser des habitats favorables à la fondation de nouvelles colonies alors que les espèces polygynes possèdent une dispersion limitée avec des accouplements à l'intérieur des nids. Afin de comparer les stratégies de reproduction de F. lugubris et de F. paralugubris (accouplements et perte des ailes) nous avons mené des expériences avec les sexués. Les jeunes femelles ailées de F. lugubris issues de populations monogynes/monodomes ne présentent pas de stratégie locale comparée à l'espèce obligatoirement polygyne F paralugubris (perte des ailes précoce même si il n'y a pas eu accouplement, insémination possible sans avoir volé activement et faibles réserves de graisse). Elles conservent toujours leurs ailes, ne s'accouplent pas lorsqu'elles sont empêchées de voler et possèdent de grandes quantités de graisse révélant de fortes capacités de survie. D'autre part, les jeunes femelles ailées de F. lugubris provenant de populations polygynes/polydomes ont peu de réserves lipidiques et ont un comportement de reproduction proche de celles de F. paralugubris. Après leur dispersion, les jeunes sexués femelles de fourmis des bois peuvent soit fonder une nouvelle société par parasitisme social temporaire d'un nid d'une autre espèce (sous-genre Serviformica) soit être adoptées dans un nid déjà existant de leur propre espèce. Chez F. lugubris, nous avons pu démontrer que les jeunes sexués femelles de colonies monogynes/monodomes présentent un succès élevé au parasitisme sociale temporaire en comparaison au plus faible succès obtenu avec des sexués provenant de colonies polygynes/polydomes. Cependant, les données physiologiques suggèrent que seules les jeunes sexués femelles de populations mono-gynes/monodomes peuvent disperser efficacement et fonder un nouveau nid par parasitisme social temporaire. Au niveau intraspécifique, les jeunes femelles sont acceptées à un taux élevé dans les nids polygynes mais aussi dans les nids monogynes tant que ces nids possèdent encore de jeunes sexués. En conclusion, F. lugubris est caractérisée par un polymorphisme dans ses structures sociales et ses stratégies de dispersion (système mixte d'accouplement et de fondation) ce qui représente une forte plasticité comportementale en relation avec les conditions environnementales et écologiques. Par conséquent, les mesures de conservation de cette espèce devraient s'attacher à maintenir un maximum de diversité au niveau des habitats.

Integrative taxonomy of the Formica rufa group (Hymenoptera: Formicidae)

RESUME :Les fourmis du groupe Formica rufa, ou fourmis des bois, ainsi appelées en raison de leur préférence pour les écosystèmes forestiers, sont parmi les fourmis les plus fascinantes et les plus étudiées d'Europe. Ces fourmis jouent un rôle clé dans la plupart des forêts dans lesquelles elles vivent et sont considérées comme étant les meilleurs bioindicateurs de ces milieux. Pour ces raisons, les fourmis des bois sont protégées par la loi dans de nombreux pays européens, y compris en Suisse. Cependant, malgré leur protection, ces fourmis sont inscrites sur la liste rouge des espèces menacées dans plusieurs pays d'Europe et il est donc indispensable de bien les connaître afin de mieux les protéger.À l'heure actuelle, on considère que le groupe Formica rufa est composé de six espèces distinctes : F. rufa, F. polyctena, F. lugubris, F. paralugubris, F. aquilonia et F. pratensis. Toutefois, malgré la grande quantité d'études effectuées sur ces espèces, la systématique et l'identification des fourmis des bois sont toujours sujettes à discussion. Ceci est essentiellement dû au fait que ces espèces sont morphologiquement similaires et qu'elles sont parfois capables de s'hybrider ou de former des colonies mixtes.Une des conditions fondamentales pour toute étude en biologie de la conservation est l'identification correcte des espèces à protéger. Avec cette étude, nous désirons donc dénouer les problèmes liés à la systématique des fourmis des bois et analyser la diversité de ces espèces en adoptant une approche multidisciplinaire.Nous avons d'abord étudié la distribution des espèces jumelles F. lugubris et F. paralugubris dans les Álpes italiennes en re-analysant l'une des plus grandes collections de références sur ces espèces, déposée à l'Université de Pavie, Italie, et en récoltant de nouveaux échantillons sur le terrain. Nos analyses ont montré que F, paralugubris, décrite récemment et souvent «oubliée »par les chercheurs, est bien présente dans les Alpes et vit souvent en sympathie avec F. lugubris. Ensuite nous avons développé un outil moléculaire basé sur l'ADN mitochondrial pour une identification rapide et efficace de ces deux espèces. Au vu des bons résultats, nous avons étendu nos analyses génétiques (microsatellites) à toutes les espèces du groupe F. rufa, ce qui nous a permis de montrer que les outils moléculaires sont très efficaces pour identifier ces fourmis. En outre, nos analyses ont mis en évidence la présence d'une nouvelle espèce cryptique (appelée F. lugubris-X) au sein du Parc National Suisse. L'existence d'une nouvelle espèce peut avoir une grande influence sur les projets de conservation en faveur de ces espèces. Nous avons donc décidé de confirmer ce résultat avec des analyses comportementales et des analyses chimiques basées sur les phéromones sexuelles des différentes espèces, y compris F. lugubris-X. Les deux approches confirment nos données génétiques et indiquent que F. lugubris-X représente bel et bien une nouvelle espèce de fourmis des bois dans les Alpes Suisses.Les résultats de cette étude ont une grande importance du point de vue de la biodiversité. En plus, ils livrent aux futurs chercheurs des outils fiables pour l'identification des fourmis des bois et ouvrent de captivantes perspectives pour une meilleure protection de ces insectes et, par conséquent, de nos écosystèmes forestiers. .Abstract :Mound building red wood ants (species of the Formica rufa group) belong to one of the most studied groups of ants in Europe and have fundamental roles and positive effects in forested habitats of the northern hemisphere. In addition, they are considered among the most promising bioindicators of forest ecosystems. Because of their importance, these ants are protected by law in many European countries, including Switzerland. However, despite this protection, they are included on the red list of threatened species edited by the International Union for Conservation of Nature (IUCN) and on the red list of some particular countries like Switzerland. Because of their similar morphology and a high intraspecific variability, the morphological identification of these species can be quite complicated. In addition, they are sometimes able to hybridize or to form mixed colonies. Consequently, the taxonomy of this group of ants has been much debated during the past decades. Based on a phylogenetic study, today the group is considered to count six species in Europe: F. rufa, F. po/yctena, F. lugubris, F. paralugubris, F. aquilonia and F. pratensis. Nevertheless, the taxonomy of the group is often neglected mainly due to the lack of reliable and easy to use identification methods.Considering the importance of correct species assessment in conservation biology, in this study we want to disentangle the taxonomical difficulties within the Formica rufa group and to clarify the diversity of these protected ants, by using an integrative approach.We first analyzed the distribution of .the sibling species F. lugubris and F. paralugubris in the Italian Alps by collecting new samples on the field and by examining one of the major red wood ant collections, which is deposited at the University of Pavia, Italy. After that, we developed a molecular tool based on mitochondria) DNA, which provides a reliable and easy-to-use technique for the identification of F. lugubris and F. paralugubris. Afterwards, we extended the use of molecular markers for species identification to the whole F. rufa group and made a microsatellite analysis. Results confirm that molecular markers are consistent tools for species identification and that the six known species represent six different genetic pools. In addition, genetic data highlighted the existence of a new cryptic species in the Swiss Alps, called Formica lugubris-X.The presence of a new species can have a great influence on future conservation plans in favour of these protected ants and consequently for forested habitats. We therefore completed molecular data by behavioural (pupae recognition) and chemical analyses based on six pheromones of the entire F. rufa group. Both approaches are in accordance to genetic results and confirm that F. lugubris-X really represents a new cryptic species of red wood ant within the Swiss National Park (Eastern Swiss Alps).Results obtained in this study have a great importance in terms of biodiversity. Moreover, they provide important taxonomical information, reliable tools for species identifications and future perspectives for a consequent conservation of red wood ant species.

The determinants of queen size in a socially polymorphic ant.

In social animals, body size can be shaped by multiple factors, such as direct genetic effects, maternal effects, or the social environment. In ants, the body size of queens correlates with the social structure of the colony: colonies headed by a single queen (monogyne) generally produce larger queens that are able to found colonies independently, whereas colonies headed by multiple queens (polygyne) tend to produce smaller queens that stay in their natal colony or disperse with workers. We performed a cross-fostering experiment to investigate the proximate causes of queen size variation in the socially polymorphic ant Formica selysi. As expected if genetic or maternal effects influence queen size, eggs originating from monogyne colonies developed into larger queens than eggs collected from polygyne colonies, be they raised by monogyne or polygyne workers. In contrast, eggs sampled in monogyne colonies were smaller than eggs sampled in polygyne colonies. Hence, eggs from monogyne colonies are smaller but develop into larger queens than eggs from polygyne colonies, independently of the social structure of the workers caring for the brood. These results demonstrate that a genetic polymorphism or maternal effect transmitted to the eggs influences queen size, which probably affects the social structure of new colonies.

Molecular substitution rate increases in myrmecophilous lycaenid butterflies (Lepidoptera)

Is species diversification driven by neutral- or niche-based processes? Butterflies of the Lycaenidae family have developed mutualistic interactions with ants. This biotic requirement increased the spatial fragmentation of populations of lower effective population size (Ne) compared with autonomous species. The nearly neutral theory predicts that species with smaller Ne should fix more mutations because of the increased strength of drift. Taking into account the phylogenetic relatedness among species, this study shows that species with a stronger dependence on ants displayed more intra-specific Single Nucleotide Polymorphisms compared with species with low or no myrmecophily. This phenomenon can cause more pronounced genetic differentiation between populations and could ultimately promote speciation in a similar manner as on physical islands. The large species diversity observed in this family could be the consequence of this neutral process enhancing the diversification of lineages.

TWEAK can induce cell death via endogenous TNF and TNF receptor 1.

TWEAK is a recently cloned novel member of the TNF ligand family. Here we show that soluble TWEAK is sufficient to induce apoptosis in Kym-1 cells within 18 h. TWEAK-induced apoptosis is indirect and is mediated by the interaction of endogenous TNF and TNF receptor (TNFR)1, as each TNFR1-Fc, neutralizing TNF-specific antibodies and TNFR1-specific Fab fragments efficiently antagonize cell death induction. In addition to this indirect mode of action, co-stimulation of Kym-1 cells with TWEAK enhances TNFR1-mediated cell death induction. In contrast to TNF, TWEAK does only modestly activate NF-kappaB or c-jun N-terminal kinase (JNK) in Kym-1 cells. Although TWEAK binding to Kym-1 cells is easily detectable by flow cytometric analysis, we found neither evidence for expression of the recently identified TWEAK receptor Apo3/TRAMP/wsl/DR3/LARD, nor indications for direct interactions of TWEAK with TNFR. Together, these characteristics of TWEAK-induced signaling in Kym-1 cells argue for the existence of an additional, still undefined non-death domain-containing TWEAK receptor in Kym-1 cells.

Foxm1 expression in prostate epithelial cells is essential for prostate carcinogenesis.

The treatment of advanced prostate cancer (PCa) remains a challenge. Identification of new molecular mechanisms that regulate PCa initiation and progression would provide targets for the development of new cancer treatments. The Foxm1 transcription factor is highly up-regulated in tumor cells, inflammatory cells, and cells of tumor microenvironment. However, its functions in different cell populations of PCa lesions are unknown. To determine the role of Foxm1 in tumor cells during PCa development, we generated two novel transgenic mouse models, one exhibiting Foxm1 gain-of-function and one exhibiting Foxm1 loss-of-function under control of the prostate epithelial-specific Probasin promoter. In the transgenic adenocarcinoma mouse prostate (TRAMP) model of PCa that uses SV40 large T antigen to induce PCa, loss of Foxm1 decreased tumor growth and metastasis. Decreased prostate tumorigenesis was associated with a decrease in tumor cell proliferation and the down-regulation of genes critical for cell proliferation and tumor metastasis, including Cdc25b, Cyclin B1, Plk-1, Lox, and Versican. In addition, tumor-associated angiogenesis was decreased, coinciding with reduced Vegf-A expression. The mRNA and protein levels of 11β-Hsd2, an enzyme playing an important role in tumor cell proliferation, were down-regulated in Foxm1-deficient PCa tumors in vivo and in Foxm1-depleted TRAMP C2 cells in vitro. Foxm1 bound to, and increased transcriptional activity of, the mouse 11β-Hsd2 promoter through the -892/-879 region, indicating that 11β-Hsd2 was a direct transcriptional target of Foxm1. Without TRAMP, overexpression of Foxm1 either alone or in combination with inhibition of a p19(ARF) tumor suppressor caused a robust epithelial hyperplasia, but was insufficient to induce progression from hyperplasia to PCa. Foxm1 expression in prostate epithelial cells is critical for prostate carcinogenesis, suggesting that inhibition of Foxm1 is a promising therapeutic approach for prostate cancer chemotherapy.

The evolution of unicoloniality in ant societies : recognition, dispersal and population genetic structure

Introduction Societies of ants, bees, wasps and termites dominate many terrestrial ecosystems (Wilson 1971). Their evolutionary and ecological success is based upon the regulation of internal conflicts (e.g. Ratnieks et al. 2006), control of diseases (e.g. Schmid-Hempel 1998) and individual skills and collective intelligence in resource acquisition, nest building and defence (e.g. Camazine 2001). Individuals in social species can pass on their genes not only directly trough their own offspring, but also indirectly by favouring the reproduction of relatives. The inclusive fitness theory of Hamilton (1963; 1964) provides a powerful explanation for the evolution of reproductive altruism and cooperation in groups with related individuals. The same theory also led to the realization that insect societies are subject to internal conflicts over reproduction. Relatedness of less-than-one is not sufficient to eliminate all incentive for individual selfishness. This would indeed require a relatedness of one, as found among cells of an organism (Hardin 1968; Keller 1999). The challenge for evolutionary biology is to understand how groups can prevent or reduce the selfish exploitation of resources by group members, and how societies with low relatedness are maintained. In social insects the evolutionary shift from single- to multiple queens colonies modified the relatedness structure, the dispersal, and the mode of colony founding (e.g. (Crozier & Pamilo 1996). In ants, the most common, and presumably ancestral mode of reproduction is the emission of winged males and females, which found a new colony independently after mating and dispersal flights (Hölldobler & Wilson 1990). The alternative reproductive tactic for ant queens in multiple-queen colonies (polygyne) is to seek to be re-accepted in their natal colonies, where they may remain as additional reproductives or subsequently disperse on foot with part of the colony (budding) (Bourke & Franks 1995; Crozier & Pamilo 1996; Hölldobler & Wilson 1990). Such ant colonies can contain up to several hundred reproductive queens with an even more numerous workforce (Cherix 1980; Cherix 1983). As a consequence in polygynous ants the relatedness among nestmates is very low, and workers raise brood of queens to which they are only distantly related (Crozier & Pamilo 1996; Queller & Strassmann 1998). Therefore workers could increase their inclusive fitness by preferentially caring for their closest relatives and discriminate against less related or foreign individuals (Keller 1997; Queller & Strassmann 2002; Tarpy et al. 2004). However, the bulk of the evidence suggests that social insects do not behave nepotistically, probably because of the costs entailed by decreased colony efficiency or discrimination errors (Keller 1997). Recently, the consensus that nepotistic behaviour does not occur in insect colonies was challenged by a study in the ant Formica fusca (Hannonen & Sundström 2003b) showing that the reproductive share of queens more closely related to workers increases during brood development. However, this pattern can be explained either by nepotism with workers preferentially rearing the brood of more closely related queens or intrinsic differences in the viability of eggs laid by queens. In the first chapter, we designed an experiment to disentangle nepotism and differences in brood viability. We tested if workers prefer to rear their kin when given the choice between highly related and unrelated brood in the ant F. exsecta. We also looked for differences in egg viability among queens and simulated if such differences in egg viability may mistakenly lead to the conclusion that workers behave nepotistically. The acceptance of queens in polygnous ants raises the question whether the varying degree of relatedness affects their share in reproduction. In such colonies workers should favour nestmate queens over foreign queens. Numerous studies have investigated reproductive skew and partitioning of reproduction among queens (Bourke et al. 1997; Fournier et al. 2004; Fournier & Keller 2001; Hammond et al. 2006; Hannonen & Sundström 2003a; Heinze et al. 2001; Kümmerli & Keller 2007; Langer et al. 2004; Pamilo & Seppä 1994; Ross 1988; Ross 1993; Rüppell et al. 2002), yet almost no information is available on whether differences among queens in their relatedness to other colony members affects their share in reproduction. Such data are necessary to compare the relative reproductive success of dispersing and non-dispersing individuals. Moreover, information on whether there is a difference in reproductive success between resident and dispersing queens is also important for our understanding of the genetic structure of ant colonies and the dynamics of within group conflicts. In chapter two, we created single-queen colonies and then introduced a foreign queens originating from another colony kept under similar conditions in order to estimate the rate of queen acceptance into foreign established colonies, and to quantify the reproductive share of resident and introduced queens. An increasing number of studies have investigated the discrimination ability between ant workers (e.g. Holzer et al. 2006; Pedersen et al. 2006), but few have addressed the recognition and discrimination behaviour of workers towards reproductive individuals entering colonies (Bennett 1988; Brown et al. 2003; Evans 1996; Fortelius et al. 1993; Kikuchi et al. 2007; Rosengren & Pamilo 1986; Stuart et al. 1993; Sundström 1997; Vásquez & Silverman in press). These studies are important, because accepting new queens will generally have a large impact on colony kin structure and inclusive fitness of workers (Heinze & Keller 2000). In chapter three, we examined whether resident workers reject young foreign queens that enter into their nest. We introduced mated queens into their natal nest, a foreign-female producing nest, or a foreign male-producing nest and measured their survival. In addition, we also introduced young virgin and mated queens into their natal nest to examine whether the mating status of the queens influences their survival and acceptance by workers. On top of polgyny, some ant species have evolved an extraordinary social organization called 'unicoloniality' (Hölldobler & Wilson 1977; Pedersen et al. 2006). In unicolonial ants, intercolony borders are absent and workers and queens mix among the physically separated nests, such that nests form one large supercolony. Super-colonies can become very large, so that direct cooperative interactions are impossible between individuals of distant nests. Unicoloniality is an evolutionary paradox and a potential problem for kin selection theory because the mixing of queens and workers between nests leads to extremely low relatedness among nestmates (Bourke & Franks 1995; Crozier & Pamilo 1996; Keller 1995). A better understanding of the evolution and maintenance of unicoloniality requests detailed information on the discrimination behavior, dispersal, population structure, and the scale of competition. Cryptic genetic population structure may provide important information on the relevant scale to be considered when measuring relatedness and the role of kin selection. Theoretical studies have shown that relatedness should be measured at the level of the `economic neighborhood', which is the scale at which intraspecific competition generally takes place (Griffin & West 2002; Kelly 1994; Queller 1994; Taylor 1992). In chapter four, we conducted alarge-scale study to determine whether the unicolonial ant Formica paralugubris forms populations that are organised in discrete supercolonies or whether there is a continuous gradation in the level of aggression that may correlate with genetic isolation by distance and/or spatial distance between nests. In chapter five, we investigated the fine-scale population structure in three populations of F. paralugubris. We have developed mitochondria) markers, which together with the nuclear markers allowed us to detect cryptic genetic clusters of nests, to obtain more precise information on the genetic differentiation within populations, and to separate male and female gene flow. These new data provide important information on the scale to be considered when measuring relatedness in native unicolonial populations.

Fourmis des bois en Suisse: état de la situation et perspectives de monitoring

The authors summarise their state of knowledge about wood ants and their role in the forest ecosystem. They also describe the situation in Switzerland, their past development and their conservation status. In several re- gions, mainly in the Plateau, wood ants seem to diminish, despite their total protection since 1966. The reasons for this regression are not well known, but the fragmentation of forest habitats in the Plateau region and direct damage to ant nests seem to play a certain role. A new project in which the development of wood ant nests is monitored in Swiss forest reserves (Formica-Forêts-CH) was recently started in the Swiss national park. It is to be extended, in collaboration with the forest services, over the whole of Switzerland.

Global invasion history of the fire ant Solenopsis invicta.

The fire ant Solenopsis invicta is a significant pest that was inadvertently introduced into the southern United States almost a century ago and more recently into California and other regions of the world. An assessment of genetic variation at a diverse set of molecular markers in 2144 fire ant colonies from 75 geographic sites worldwide revealed that at least nine separate introductions of S. invicta have occurred into newly invaded areas and that the main southern U.S. population is probably the source of all but one of these introductions. The sole exception involves a putative serial invasion from the southern United States to California to Taiwan. These results illustrate in stark fashion a severe negative consequence of an increasingly massive and interconnected global trade and travel system.

Note préliminaire sur la structure, la phénologie et le régime alimentaire d'une super-colonie de formica lugubris zett

Formica lugubris apparaît comme une espèce hautement polycalique dans le Jura suisse et forme des super-colonies. La super-colonie étudiée comprend environ 1200 nids répartis sur 70 hectares. L'étude détaillée de 12 hectares permet de définir 4 types de nids:les nids principaux, secondaires, saisonniers etcommençants, ainsi que trois sortes de voies de communication:les routes de liaisons permanentes visibles sur le terrain, les pistes de liaisons non-permanentes non marquées sur le terrain etles chemins d'approvisionnement permanents marqués dans le terrain. L'auteur présente la phénologie deF. lugubris qui est fortement influencée par le climat de cette région avec une période moyenne d'activité de 150 jours. D'autre part, les premières données sur le régime alimentaire (analyse des proies récoltées par les fourmis) diffèrent considérablement des données connues pour les autres espèces du groupe rufa, notamment par le nombre élevé de pucerons, d'où l'idée d'une régulation des populations de pucerons par les fourmis. Enfin l'auteur aborde le problème de la faible densité de l'avifaune en relation avec les fourmis. Il semble que le climat et les ressources alimentaires conduisent les fourmis àune nouvelle stratégie écologique qui s'exprimerait par la création de super-colonies. Formica lugubris appears as a highly polycalic species in the Swiss Jura and creates super-colonies. The super-colony studied possesses about 1200 nests on about 70 hectares. The detailed study of 12 hectares allows the discrimination of 4 types of nests:the main nests, the secondary nests, the seasonal nests andthe starting nests, as well as 3 types of ant tracks:the constant connection routes visible on the soil, thenon-constant connection tracks not marked on the soil andthe constant foraging routes marked on the soil. The author presents the phenology ofF. lugubris who is strongly influenced by the climate of the region with a mean activity period of about 150 days. On the other hand, the first results about diet (analysis of the preys collected by the ants) differ considerably from the wellknown data for the others species of the rufa group, especially by the high number of aphids, which may be inferred the notion of a regulation of aphids population by the ants. Finally the author approaches the problem of the low density of avifauna in relation to the ants. It seems that climate and food resources lead the ants toa new ecological strategy which would express itself by the creation of super-colonies.

Variable queen number in ant colonies: no impact on queen turnover, inbreeding, and population genetic differentiation in the ant Formica selysi.

Variation in queen number alters the genetic structure of social insect colonies, which in turn affects patterns of kin-selected conflict and cooperation. Theory suggests that shifts from single- to multiple-queen colonies are often associated with other changes in the breeding system, such as higher queen turnover, more local mating, and restricted dispersal. These changes may restrict gene flow between the two types of colonies and it has been suggested that this might ultimately lead to sympatric speciation. We performed a detailed microsatellite analysis of a large population of the ant Formica selysi, which revealed extensive variation in social structure, with 71 colonies headed by a single queen and 41 by multiple queens. This polymorphism in social structure appeared stable over time, since little change in the number of queens per colony was detected over a five-year period. Apart from queen number, single- and multiple-queen colonies had very similar breeding systems. Queen turnover was absent or very low in both types of colonies. Single- and multiple-queen colonies exhibited very small but significant levels of inbreeding, which indicates a slight deviation from random mating at a local scale and suggests that a small proportion of queens mate with related males. For both types of colonies, there was very little genetic structuring above the level of the nest, with no sign of isolation by distance. These similarities in the breeding systems were associated with a complete lack of genetic differentiation between single- and multiple-queen colonies, which provides no support for the hypothesis that change in queen number leads to restricted gene flow between social forms. Overall, this study suggests that the higher rates of queen turnover, local mating, and population structuring that are often associated with multiple-queen colonies do not appear when single- and multiple-queen colonies still coexist within the same population, but build up over time in populations consisting mostly of multiple-queen colonies.

Population genetic structure in the native and introduced range of the Argentine ant/

SUMMARY : The evolution of animal societies, where some individuals forego their own reproductive opportunities to help others to reproduce, poses an evolutionary paradox that can be traced back to Darwin. Altruism may evolve through kin selection when the donor and recipient of altruistic acts are related to each other. In social insects, workers are generally highly related to the brood they rear when colonies are headed by a single queen. Yet some ants have an extraordinary social organization, called unicoloniality, whereby individuals from separate nests mix freely to form large supercolonies, which in some cases extend over hundreds of km. These supercolonies are characterised by a high number of queens (polygyny) and an absence of clear colony boundaries. This type of social organization represents an evolutionary paradox because relatedness between nestmates is effectively zero. In such conditions, kin selection cannot account for the evolution of reproductive altruism. Moreover, unicoloniality is thought to be unstable over time, because workers that can no longer aid close relatives may evolve more selfish strategies. The Argentine ant (Linepithema humile) is a highly invasive species listed among the hundred world's worst invaders by the UICN. Native from South America, L. humile has been accidentally introduced throughout the world. Native populations have been described as noninvasive with a family-based organization. In contrast, within its introduction range, they form unicolonial supercolonies that contain numerous nests without intraspecific aggression. The development of such unicolonial populations has been explained as a direct consequence of the ant's introduction into a new habitat, favouring a transition from family-based to open colonies. To determine if the social structure of the Argentine ant is fundamentally different between the native and the introduced range, we studied genetically and behaviourally native and introduced populations of L. humile over different geographic scales. Our results clearly indicated that there are no fundamental differences in the social organisation of the Argentine ant between the two ranges. Our investigations revealed that, contrary to previous claims, native populations have a unicolonial social organisation very similar to that observed in the introduced range. Consequently, the unicolonial social structure of the Argentine ant does not stem from a shift in social organization associated with introduction into new habitats but evolved in the native range and is likely a stable, evolutionarily ancient adaptation to the local environment. Our study on native populations of L. humile also gave important insight in the comprehension of the evolution of unicoloniality in the Argentine ant. Native supercolonies are relatively small compared to introduced ones and may co-habit in a same population. These supercolonies are genetically highly differentiated leading to a significant relatedness among nestmate workers when the different supercolonies of a population are taken as a reference population. This provides the necessary conditions for loin selection to operate. Furthermore, we examined a native population over time, which revealed a high supercolony extinction rate. If more competitive supercolonies are more likely to survive or replace other supercolonies, a subtle dynamical process between the spread of selfish traits within supercolony and the selective elimination of supercolonies with such traits may allow a stable equilibrium and the persistence of unicoloniality over time. Finally, a worldwide study of the Argentine ant showed that the introduced supercolonies originate from numerous independent introduction events. In conclusion, the success of the Argentine ant does not stem from a shift in social organization associated with its introduction into new habitats, but is most probably explained by the intrinsic characteristics developed in its native range. RESUME : L'altruisme de reproduction où certains individus renoncent à leur propre reproduction pour aider d'autres individus à se reproduire constitue l'un des plus grand paradoxe de l'évolution. En effet, comment expliquer l'évolution de comportements qui tendent à augmenter les chances de survie et le succès reproductif d'autres individus, alors que ces actes diminuent l'aptitude de leurs auteurs ? La théorie de la sélection de parentèle permet de résoudre ce problème. Cette théorie stipule qu'en aidant de proches parents à se reproduire, les individus peuvent promouvoir indirectement la transmission de copies de leurs propres gènes à la génération suivante. Chez les insectes sociaux, l'altruisme des ouvrières s'explique par la théorie de sélection de parentèle lorsque les colonies sont monogynes (constituées d'une seule reine) puisque les ouvrières sont fortement apparentées aux couvains dont elles s'occupent. Par contre, les espèces dites unicoloniales, dont les colonies forment des réseaux de nids appelés supercolonies, représentent toujours un paradoxe pour les théories de l'évolution puisque l'apparentement entre les différents individus d'un nid est nulle. De plus, l'unicolonialité ne devrait pas être stable sur le long terme parce que les ouvrières qui ne s'occupent plus de leur apparentés devraient développer des stratégies plus égoïstes au cours du temps. La fourmi d'Argentine (Linepithema humile) est une espèce invasive ayant un impact considérable sur son environnement. Originaire d'Amérique du Sud, elle a été introduite dans pratiquement toutes les régions du monde dont le climat est de type méditerranéen. Son incroyable succès invasif s'explique par sa structure sociale unicoloniale observée dans chacun des pays où elle a été introduite. Par contre, les rares études effectuées en Argentine ont suggéré que la fourmi d'Argentine n'était pas unicoloniale dans son aire native. L'unicolonialité chez la fourmi d'Argentine était donc considéré comme une conséquence de son introduction dans de nouveaux environnements. Durant cette thèse, nous avons vérifié si la structure sociale de cette espèce différait fondamentalement entre l'aire native et introduite. Pour cela, nous avons étudié, à différentes échelles géographiques, des populations introduites et argentines avec une approche génétique et comportementale. L'ensemble de nos résultats montrent que les différences entre les deux structure sociales ne sont pas aussi importantes que ce que l'on imaginait. Les populations natives sont aussi constituées de réseaux de nids coopérants. La taille de ses supercolonies est toutefois bien moins importante en Argentine et il n'est pas rare de trouver plusieurs supercolonies cohabitantes dans une même population. Nous avons démontré que ces réseaux de nids étaient constitués d'individus qui sont plus apparentés entre eux qu'ils ne le sont avec les individus d'autres supercolonies, ainsi l'unicolonialité dans son aire d'origine ne représente pas un réel paradoxe pour les théories de l'évolution. Finalement nous avons étudié la même population en Argentine à six ans d'intervalle et avons constaté que les supercolonies avaient un taux de survie très faible ce qui pourrait expliquer la stabilité de l'unicolonialité au cours du temps. Si les supercolonies les plus compétitives survivent mieux que les supercolonies dans lesquelles apparaissent des traits égoïstes, on devrait alors observer une dynamique entre l'apparition de traits égoïstes et l'élimination des supercolonies dans lesquelles ces traits égoïstes évolueraient. Finalement, une étude mondiale nous a montré que les supercolonies étaient originaires de nombreux événements d'introductions indépendants. En conclusion, le succès invasif de la fourmi d'Argentine n'est donc pas dû à un changement de comportement associé à son introduction mais est lié aux caractéristiques qu'elle a développées en Argentine.

Unicoloniality, recognition and genetic differentiation in a native Formica ant.

Some ants have an extraordinary form of social organization, called unicoloniality, whereby individuals mix freely among physically separated nests. This mode of social organization has been primarily studied in introduced and invasive ant species, so that the recognition ability and genetic structure of ants forming unicolonial populations in their native range remain poorly known. We investigated the pattern of aggression and the genetic structure of six unicolonial populations of the ant Formica paralugubris at four hierarchical levels: within nests, among nests within the same population, among nests of populations within the Alps or Jura Mountains and among nests of the two mountain ranges. Ants within populations showed no aggressive behaviour, but recognized nonnestmates as shown by longer antennation bouts. Overall, the level of aggression increased with geographic and genetic distance but was always considerably lower than between species. No distinct behavioural supercolony boundaries were found. Our study provides evidence that unicoloniality can be maintained in noninvasive ants despite significant genetic differentiation and the ability to discriminate between nestmates and nonnestmates.

Odorant binding proteins of the red imported fire ant, Solenopsis invicta: an example of the problems facing the analysis of widely divergent proteins.

We describe the odorant binding proteins (OBPs) of the red imported fire ant, Solenopsis invicta, obtained from analyses of an EST library and separate 454 sequencing runs of two normalized cDNA libraries. We identified a total of 18 putative functional OBPs in this ant. A third of the fire ant OBPs are orthologs to honey bee OBPs. Another third of the OBPs belong to a lineage-specific expansion, which is a common feature of insect OBP evolution. Like other OBPs, the different fire ant OBPs share little sequence similarity (∼ 20%), rendering evolutionary analyses difficult. We discuss the resulting problems with sequence alignment, phylogenetic analysis, and tests of selection. As previously suggested, our results underscore the importance for careful exploration of the sensitivity to the effects of alignment methods for data comprising widely divergent sequences.