British Red Squirrels Remain the Only Known Wild Rodent Host for Leprosy Bacilli.

Eurasian red squirrels (Sciurus vulgaris) in the British Isles are the most recently discovered animal reservoir for the leprosy bacteria Mycobacterium leprae and Mycobacterium lepromatosis. Initial data suggest that prevalence of leprosy infection is variable and often low in different squirrel populations. Nothing is known about the presence of leprosy bacilli in other wild squirrel species despite two others (Siberian chipmunk [Tamias sibiricus], and Thirteen-lined ground squirrel [Ictidomys tridecemlineatus]) having been reported to be susceptible to experimental infection with M. leprae. Rats, a food-source in some countries where human leprosy occurs, have been suggested as potential reservoirs for leprosy bacilli, but no evidence supporting this hypothesis is currently available. We screened 301 squirrel samples covering four species [96 Eurasian red squirrels, 67 Eastern gray squirrels (Sciurus carolinensis), 35 Siberian chipmunks, and 103 Pallas's squirrels (Callosciurus erythraeus)] from Europe and 72 Mexican white-throated woodrats (Neotoma albigula) for the presence of M. leprae and M. lepromatosis using validated PCR protocols. No DNA from leprosy bacilli was detected in any of the samples tested. Given our sample-size, the pathogen should have been detected if the prevalence and/or bacillary load in the populations investigated were similar to those found for British red squirrels.

Within- and between-year variations of reproductive strategy and cost in a population of Siberian chipmunks.

Reproduction costs depend on the general life-history strategies employed by organisms for resource acquisition, the decision rules on resource allocation, and the resource availability. Although the predictability of resource availability is expected to influence the breeding strategy, the relationship between predictability and strategy has rarely been investigated at the population level. One reason is that, while the resource availability is commonly variable in space and time, their predictability is generally assumed constant. Here, we addressed the temporal variation of the breeding strategy and its associated survival cost in a hibernating population of Tamias sibiricus, in which food resources vary in their availability between years and in their predictability within years. Based on 11 years of mark-recapture data, we used multi-event modelling to investigate seasonal variations in reproduction costs of female chipmunks that breed twice a year (spring and summer). In summer, during which a large variety and quantity of resources is available (income breeding strategy), the proportion of breeding females was consistent across years and reproduction yielded no mortality cost. In contrast, in spring, the proportion of breeding females was positively correlated with the amount of resources available for hibernation (partial capital breeding strategy). Spring reproduction yielded no immediate cost, but induced a delayed mortality cost over the next winter if future unknown conditions were unfavorable. Our findings highlight complex temporal reproductive patterns in a short-lived species: not only does the modality of resource acquisition vary among seasons, but also the decision rule to breed and its associated cost.

Divergent in shape and convergent in function: Adaptive evolution of the mandible in Sub-Antarctic mice.

Convergent evolution in similar environments constitutes strong evidence of adaptive evolution. Transported with people around the world, house mice colonized even remote areas, such as Sub-Antarctic islands. There, they returned to a feral way of life, shifting towards a diet enriched in terrestrial macroinvertebrates. Here, we test the hypothesis that this triggered convergent evolution of the mandible, a morphological character involved in food consumption. Mandible shape from four Sub-Antarctic islands was compared to phylogeny, tracing the history of colonization, and climatic conditions. Mandible shape was primarily influenced by phylogenetic history, thus discarding the hypothesis of convergent evolution. The biomechanical properties of the jaw were then investigated. Incisor in-lever and temporalis out-lever suggested an increase in the velocity of incisor biting, in agreement with observations on various carnivorous and insectivorous rodents. The mechanical advantage related to incisor biting also revealed an increased functional performance in Sub-Antarctic populations, and appears to be an adaptation to catch prey more efficiently. The amount of change involved was larger than expected for a plastic response, suggesting microevolutionary processes were evolved. This study thus denotes some degree of adaptive convergent evolution related to changes in habitat-related changes in dietary items in Sub-Antarctic mice, but only regarding simple, functionally relevant aspects of mandible morphology.

Multiple independent transmission cycles of a tick-borne pathogen within a local host community.

Many pathogens are maintained by multiple host species and involve multiple strains with potentially different phenotypic characteristics. Disentangling transmission patterns in such systems is often challenging, yet investigating how different host species contribute to transmission is crucial to properly assess and manage disease risk. We aim to reveal transmission cycles of bacteria within the Borrelia burgdorferi species complex, which include Lyme disease agents. We characterized Borrelia genotypes found in 488 infected Ixodes ricinus nymphs collected in the Sénart Forest located near Paris (France). These genotypes were compared to those observed in three sympatric species of small mammals and network analyses reveal four independent transmission cycles. Statistical modelling shows that two cycles involving chipmunks, an introduced species, and non-sampled host species such as birds, are responsible for the majority of tick infections. In contrast, the cycle involving native bank voles only accounts for a small proportion of infected ticks. Genotypes associated with the two primary transmission cycles were isolated from Lyme disease patients, confirming the epidemiological threat posed by these strains. Our work demonstrates that combining high-throughput sequence typing with networks tools and statistical modeling is a promising approach for characterizing transmission cycles of multi-host pathogens in complex ecological settings.

Inter-annual and inter-individual variations in survival exhibit strong seasonality in a hibernating rodent.

Most research on the demography of wild animal populations has focused on characterizing the variation in the mortality of organisms as a function of intrinsic and environmental characteristics. However, such variation in mortality is difficult to relate to functional life history components (e.g. reproduction, dispersal, hibernation) due to the difficulty in monitoring biological processes at a sufficiently fine timescale. In this study, we used a 10-year individual-based data set with an infra-annual timescale to investigate both intra- and inter-annual survival patterns according to intrinsic and environmental covariates in an introduced population of a small hibernating rodent, the Siberian chipmunk. We compared three distinct periods related to particular life history events: spring reproduction, summer reproduction and hibernation. Our results revealed strong interactions between intrinsic and temporal effects. First, survival of male chipmunks strongly decreases during the reproduction periods, while survival is high and equal between sexes during hibernation. Second, the season of birth affects the survival of juveniles during their first hibernation, which does not have long-lasting consequences at the adult stage. Third, at an inter-annual scale, we found that high food resource availability before hibernation and low chipmunk densities specifically favour subsequent winter survival. Overall, our results confirm that the well-known patterns of yearly and inter-individual variation of mortality observed in animals are themselves strongly variable within a given year, suggesting that they are associated with various functional components of the animals' life history.

Phylogeny and adaptation shape the teeth of insular mice.

By accompanying human travels since prehistorical times, the house mouse dispersed widely throughout the world, and colonized many islands. The origin of the travellers determined the phylogenetic source of the insular mice, which encountered diverse ecological and environmental conditions on the various islands. Insular mice are thus an exceptional model to disentangle the relative role of phylogeny, ecology and climate in evolution. Molar shape is known to vary according to phylogeny and to respond to adaptation. Using for the first time a three-dimensional geometric morphometric approach, compared with a classical two-dimensional quantification, the relative effects of size variation, phylogeny, climate and ecology were investigated on molar shape diversity across a variety of islands. Phylogeny emerged as the factor of prime importance in shaping the molar. Changes in competition level, mostly driven by the presence or absence of the wood mouse on the different islands, appeared as the second most important effect. Climate and size differences accounted for slight shape variation. This evidences a balanced role of random differentiation related to history of colonization, and of adaptation possibly related to resource exploitation.

Preventing species invasion: A role for integrative taxonomy?

Integrative taxonomy, a multi-disciplinary approach adding modern techniques to traditional morphology-based methods (e.g. molecular and morphological criteria), can play an important role in bioinvasion research to identify introduced taxa, discover pathways of introduction and inform authorities to control and prevent future introductions. The present study is the first on introduced populations of Callosciurus, Asiatic tree squirrels, known as potentially invasive species in Europe (Italy, Belgium and France). We combined molecular (mitochondrial DNA markers: CoxI, D-loop) and morphometric analysis on skulls, comparing them to the widest morphological and molecular datasets ever assembled for Callosciurus. Squirrels collected in Italy and Belgium share the same haplotypes and skull characteristics, but are conspicuously different from the French population in Antibes. Genetic data revealed close similarity between French squirrels and Pallas's squirrels, Callosciurus erythraeus, from Taiwan, China. Italian and Belgian squirrels formed an independent taxonomic lineage in genetic analyses, whose taxonomic rank needs further investigation. The morphological and morphometric characteristics of these 2 populations are, however, similar to known specimens assigned to Callosciurus erythraeus. These results may indicate a common origin for the populations found in Belgium and Italy. In contrast, French specimens suggest an independent introduction event of squirrels originating from Asia.

Infection of Siberian chipmunks (Tamias sibiricus barberi) with Borrelia sp. reveals a low reservoir competence under experimental conditions.

Reservoir competence is a key parameter in understanding the role of host species in the epidemiology of multi-host-especially vector-borne-pathogens. With this aim in view, we studied the reservoir competence of the Siberian chipmunk (Tamias sibiricus barberi) recently introduced into Europe, for the multi-host tick-borne bacteria, Borrelia burgdorferi sl, the agent of Lyme borreliosis. T. sibiricus were experimentally exposed to bites from Ixodes ricinus ticks infected with Borrelia burgdorferi sensu stricto and Borrelia afzelii, with subsequent assessment of bacteremia and antibody responses. Borrelia was detected in chipmunk blood samples, ear biopsies and organ necropsies, and in nymphs used for xenodiagnosis (at one and six months after the initial chipmunk infection) via both serological and molecular methods. In total, eight out of twelve chipmunks showed evidence of infection by Borrelia sp., either by ELISA or PCR. Five chipmunks developed an immune response against the bacteria one month after infection. Borrelia infection in at least one organ was observed in seven animals at 14, 38, 93 or 178 days post-infection. Xenodiagnosis was positive for one chipmunk at 38 days, but no longer at 178 days post-infection. Four chipmunks remained uninfected, despite similar infection pressures to those observed in the field. Taken together, these results suggest that chipmunks can be infected through Borrelia-infected tick bites, and can transmit Borrelia to nymphs, but do not remain persistently infected.

Seasonal variation in infestations by ixodids on Siberian chipmunks: effects of host age, sex, and birth season.

In mammalian hosts, macroparasite aggregation is highly heterogeneous over space and time and among individuals. While the exact causes of this heterogeneity remain unclear, it has mainly been attributed to individual differences in exposure and susceptibility. Although some extrinsic (e.g., parasite availability) and intrinsic (e.g., sex or age) factors are well known to affect infestation patterns, the joint and possibly interacting effects of these factors are poorly understood. Here, we study the infestation of hard ticks (mainly Ixodes ricinus) in a small rodent, the Siberian chipmunk (Tamias sibiricus), introduced to an oak-hornbeam forest in France. We investigate the seasonal variation in infestation according to the sex, age, birth season (spring-born or summer-born), and body weight of individual hosts while controlling for interannual variability. During the 10-year study period, 3421 tick count events were recorded involving 1017 chipmunks monitored by the capture-mark-recapture procedure. Our results reveal a male-biased parasitism in the Siberian chipmunk, which is not consistent among individuals born in different seasons. This sex bias is observed among spring-born juveniles from July to the beginning of hibernation. For adults, this difference becomes apparent along the reproduction period (May-September) for summer-born adults only. These complex interactions between sex, age, and birth season suggest overall that the seasonal variation of tick load is critically linked to the reproductive behavior of this small ground sciurid.

A viable population of the European red squirrel in an urban park.

Whether urban parks can maintain viable and self-sustaining populations over the long term is questionable. In highly urbanized landscapes, urban parks could play a role in biodiversity conservation by providing habitat and resources to native species. However, populations inhabiting urban parks are usually small and isolated, leading to increased demographic stochasticity and genetic drift, with expected negative consequences on their viability. Here, we investigated a European red squirrel population located in an urban park close to Paris, France (Parc de Sceaux; 184 ha) to assess its viability. Using mitochondrial D-loop sequences and 13 microsatellite loci, we showed that the population presented high levels of genetic variation and no evidence of inbreeding. The size of the population was estimated at 100-120 individuals based on the comparison of two census techniques, Distance Sampling and Capture-Mark-Recapture. The estimated heterozygosity level and population size were integrated in a Population Viability Analysis to project the likelihood of the population's persistence over time. Results indicate that the red squirrel population of this urban park can be viable on the long term (i.e. 20 years) for a range of realistic demographic parameters (juvenile survival at least >40%) and immigration rates (at least one immigration event every two years). This study highlights that urban parks can be potential suitable refuges for the red squirrel, a locally threatened species across western European countries, provided that ecological corridors are maintained.

High-throughput sequence typing reveals genetic differentiation and host specialization among populations of the Borrelia burgdorferi species complex that infect rodents.

Lyme disease is a zoonosis caused by various species belonging to the Borrelia burgdorferi bacterial species complex. These pathogens are transmitted by ticks and infect multiple, taxonomically distinct, host species. From an epidemiological perspective, it is important to determine whether genetic variants within the species complex are able to spread freely through the whole host community or, instead, if certain variants are restricted to particular hosts. To this end, we characterized the genotypes of members of the B. burgdorferi species complex; the bacteria were isolated from more than two hundred individuals captured in the wild and belonging to three different rodent host species. For each individual, we used a high-throughput approach to amplify and sequence rplB, a housekeeping gene, and ospC, which is involved in infection. This approach allowed us to evaluate the genetic diversity both within and among species in the B. burgdorferi species complex. Strong evidence of genetic differentiation among host species was revealed by both genes, even though they are, a priori, not constrained by the same selective pressures. These data are discussed in the context of the advancements made possible by multi-locus high-throughput sequencing and current knowledge of Lyme disease epidemiology.

High prevalence of Borrelia burgdorferi s.l. in the European red squirrel Sciurus vulgaris in France.

The European red squirrel (Sciurus vulgaris) has long been suspected to be a reservoir host of the agents of Lyme borreliosis, in particular B. burgdorferi sensu stricto (s.s.). However, very few data support this hypothesis. Hereafter, we investigated the infections with B. burgdorferi genospecies in road-killed red squirrels collected across France. We also characterized the diversity of hard tick species collected from a subsample of hosts. DNA of B. burgdorferi genospecies were detected and identified from PCR products in ear biopsies using reverse line blot hybridization. Variation in prevalence was investigated accord-ing to biogeographic areas (Mediterranean, Atlantic, Continental, and Alpine), season, sex, relative age, and body mass from 273 squirrels collected 2003­2010. Among the 746 identified tick specimens, no adult was observed, 63% were nymphs, and 37% were larvae all belonging to the species Ixodes ricinus except one nymph identified as I. trianguliceps. Overall, no squirrels of Mediterranean origin and no unweaned juveniles were found infested by hard ticks. Only season explained variation in I. ricinus abundance on squirrels, with more ticks present in spring to summer than in autumn to winter. Squirrels of Mediterranean origin (n = 20) were not found infected with B. burgdorferi sensu-lato (s.l.), which is almost certainly related to the low occurrence of I. ricinus in this region. Based on individuals analyzed in the other regions of France, 11.5% (n = 26) unweaned young harboured B. burgdorferi s.l., which indicates that infection occurred already in the nest. In adults (n = 227), the prevalence of infection with B. burgdorferi s.l. was 27.3%, with 18.9% B. burgdorferi s.s., 11.9% B. afzelii, and 3.5% B. garinii. The season and the body mass, sex, and geographic origin of adults had no effect on the frequency of infection. Infection prevalence of S. vulgaris is among the highest found in rodents in Europe, particularly for B. burgdorferi s.s. supporting the hypothesis that sciurids are particularly suitable hosts for this genospecies.

A probabilistic model in cross-sectional studies for identifying interactions between two persistent vector-borne pathogens in reservoir populations.

BACKGROUND: In natural populations, individuals are infected more often by several pathogens than by just one. In such a context, pathogens can interact. This interaction could modify the probability of infection by subsequent pathogens. Identifying when pathogen associations correspond to biological interactions is a challenge in cross-sectional studies where the sequence of infection cannot be demonstrated. METHODOLOGY/PRINCIPAL FINDINGS: Here we modelled the probability of an individual being infected by one and then another pathogen, using a probabilistic model and maximum likelihood statistics. Our model was developed to apply to cross-sectional data, vector-borne and persistent pathogens, and to take into account confounding factors. Our modelling approach was more powerful than the commonly used Chi-square test of independence. Our model was applied to detect potential interaction between Borrelia afzelii and Bartonella spp. that infected a bank vole population at 11% and 57% respectively. No interaction was identified. CONCLUSIONS/SIGNIFICANCE: The modelling approach we proposed is powerful and can identify the direction of potential interaction. Such an approach can be adapted to other types of pathogens, such as non-persistents. The model can be used to identify when co-occurrence patterns correspond to pathogen interactions, which will contribute to understanding how organism communities are assembled and structured. In the long term, the model's capacity to better identify pathogen interactions will improve understanding of infectious risk.

Deciphering bartonella diversity, recombination, and host specificity in a rodent community.

Host-specificity is an intrinsic feature of many bacterial pathogens, resulting from a long history of co-adaptation between bacteria and their hosts. Alpha-proteobacteria belonging to the genus Bartonella infect the erythrocytes of a wide range of mammal orders, including rodents. In this study, we performed genetic analysis of Bartonella colonizing a rodent community dominated by bank voles (Myodes glareolus) and wood mice (Apodemus sylvaticus) in a French suburban forest to evaluate their diversity, their capacity to recombine and their level of host specificity. Following the analysis of 550 rodents, we detected 63 distinct genotypes related to B. taylorii, B. grahamii, B. doshiae and a new B. rochalimae-like species. Investigating the most highly represented species, we showed that B. taylorii strain diversity was markedly higher than that of B. grahamii, suggesting a possible severe bottleneck for the latter species. The majority of recovered genotypes presented a strong association with either bank voles or wood mice, with the exception of three B. taylorii genotypes which had a broader host range. Despite the physical barriers created by host specificity, we observed lateral gene transfer between Bartonella genotypes associated with wood mice and Bartonella adapted to bank voles, suggesting that those genotypes might co-habit during their life cycle.

Macroparasite community of the Eurasian red squirrel (Sciurus vulgaris): poor species richness and diversity.

The Eurasian red squirrel (Sciurus vulgaris) is the only naturally occurring tree squirrel throughout its range. We aim at improving current knowledge on its macroparasite fauna, expecting that it will have a poor parasite diversity because in species that have no sympatric congeners parasite richness should be lower than in hosts sharing their range with several closely related species, where host-switching events and lateral transmission are promoted. We examined gastro-intestinal helminth and ectoparasite communities (excluding mites) of, respectively, 147 and 311 red squirrel roadkills collected in four biogeographic regions in Italy and France. As expected, the macroparasite fauna was poor: we found five species of nematodes and some unidentified cestodes, three fleas, two sucking lice and two hard ticks. The helminth community was dominated by a single species, the oxyurid Trypanoxyuris (Rodentoxyuris) sciuri (prevalence, 87%; mean abundance, 373 ± 65 worms/host). Its abundance varied among seasons and biogeographic regions and increased with body mass in male hosts while decreased in females. The most prevalent ectoparasites were the flea Ceratophyllus (Monopsyllus) sciurorum (28%), whose presence was affected by season, and the generalist tick Ixodes (Ixodes) ricinus that was found only in France (34%). All the other helminths and arthropod species were rare, with prevalence below 10%. However, the first record of Strongyloides robustus, a common nematode of North American Eastern grey squirrels (S. carolinensis), in two red squirrels living in areas where this alien species co-inhabits, deserves further attention, since low parasite richness could result in native red squirrels being particularly vulnerable to parasite spillover.

Introduced Siberian chipmunks (Tamias sibiricus barberi) contribute more to lyme borreliosis risk than native reservoir rodents.

The variation of the composition in species of host communities can modify the risk of disease transmission. In particular, the introduction of a new host species can increase health threats by adding a new reservoir and/or by amplifying the circulation of either exotic or native pathogens. Lyme borreliosis is a multi-host vector-borne disease caused by bacteria belonging to the Borrelia burgdorferi sensu lato complex. It is transmitted by the bite of hard ticks, especially Ixodes ricinus in Europe. Previous studies showed that the Siberian chipmunk, Tamias sibiricus barberi, an introduced ground squirrel in the Forest of Sénart (near Paris, France) was highly infested by I. ricinus, and consequently infected by B. burgdorferi sl. An index of the contribution of chipmunks to the density of infected questing nymphs on the vegetation (i.e., the acarological risk for humans) was compared to that of bank voles (Myodes glareolus) and of wood mice (Apodemus sylvaticus), two known native and sympatric competent reservoir hosts. Chipmunks produced nearly 8.5 times more infected questing nymphs than voles and mice. Furthermore, they contribute to a higher diversity of B. burgdorferi sl genospecies (B. afzelii, B. burgdorferi sensu stricto and B. garinii). The contribution of chipmunks varied between years and seasons, according to tick availability. As T. s. barberi must be a competent reservoir, it should amplify B. burgdorferi sl infection, hence increasing the risk of Lyme borreliosis in humans.

Genetic structure of the French red squirrel populations: implication for conservation.

The decline of the red squirrel (Sciurus vulgaris) in several European countries due to the introduction of the American grey squirrel (S. carolinensis) and the predicted arrival of the grey squirrel in France in the near future has lead to the development of a preventative conservation project in this country. In this study, we conducted an extensive survey of mitochondrial DNA variation in French red squirrels using a fragment of the mitochondrial D-loop and we compared the results with previously published data from other European populations. Our main aims were: (1) to determine whether genetically differentiated populations, which could represent prioritized units for conservation purposes, were present in France and (2) to determine whether the French population, which is currently largely undisturbed, could provide information on the postglacial recolonization history of the species. We found that French D-loop haplotypes show almost no tendency to cluster by geographic origin, be it region or country, suggesting that French red squirrels have not been isolated from other populations during an evolutionarily significant period and that they do not constitute an Evolutionary Significant Unit. The French red squirrels showed strong signals of population expansion, the opposite to what is observed in most other European populations, making them of particular interest to study the postglacial expansion history of the species.

Co-infection of Borrelia afzelii and Bartonella spp. in bank voles from a suburban forest.

We report the molecular detection of Borrelia afzelii (11%) and Bartonella spp. (56%) in 447 bank voles trapped in a suburban forest in France. Adult voles were infected by significantly more Borrelia afzelii than juveniles (p<0.001), whereas no significant difference was detected in the prevalence of Bartonella spp. between young and adult individuals (p=0.914). Six percent of the animals were co-infected by both bacteria. Analysis of the bank vole carrier status for either pathogen indicated that co-infections occur randomly (p=0.94, CI(95)=[0.53; 1.47]). Sequence analysis revealed that bank voles were infected by a single genotype of Borrelia afzelii and by 32 different Bartonella spp. genotypes, related to three known species specific to rodents (B. taylorii, B. grahamii and B. doshiae) and also two as yet unidentified Bartonella species. Our findings confirm that rodents harbor high levels of potential human pathogens; therefore, widespread surveillance should be undertaken in areas where humans may encounter rodents.

Seabird modulations of isotopic nitrogen on islands.

The transport of nutrients by migratory animals across ecosystem boundaries can significantly enrich recipient food webs, thereby shaping the ecosystems' structure and function. To illustrate the potential role of islands in enabling the transfer of matter across ecosystem boundaries to be gauged, we investigated the influence of seabirds on nitrogen input on islands. Basing our study on four widely differing islands in terms of their biogeography and ecological characteristics, sampled at different spatial and temporal intervals, we analyzed the nitrogen isotopic values of the main terrestrial ecosystem compartments (vascular plants, arthropods, lizards and rodents) and their relationship to seabird values. For each island, the isotopic values of the ecosystem were driven by those of seabirds, which ultimately corresponded to changes in their marine prey. First, terrestrial compartments sampled within seabird colonies were the most enriched in δ(15)N compared with those collected at various distances outside colonies. Second, isotopic values of the whole terrestrial ecosystems changed over time, reflecting the values of seabirds and their prey, showing a fast turnover throughout the ecosystems. Our results demonstrate that seabird-derived nutrients not only spread across the terrestrial ecosystems and trophic webs, but also modulate their isotopic values locally and temporally on these islands. The wealth of experimental possibilities in insular ecosystems justifies greater use of these model systems to further our understanding of the modalities of trans-boundary nutrient transfers.

House mouse colonization patterns on the sub-Antarctic Kerguelen Archipelago suggest singular primary invasions and resilience against re-invasion.

BACKGROUND: Starting from Western Europe, the house mouse (Mus musculus domesticus) has spread across the globe in historic times. However, most oceanic islands were colonized by mice only within the past 300 years. This makes them an excellent model for studying the evolutionary processes during early stages of new colonization. We have focused here on the Kerguelen Archipelago, located within the sub-Antarctic area and compare the patterns with samples from other Southern Ocean islands. RESULTS: We have typed 18 autosomal and six Y-chromosomal microsatellite loci and obtained mitochondrial D-loop sequences for a total of 534 samples, mainly from the Kerguelen Archipelago, but also from the Falkland Islands, Marion Island, Amsterdam Island, Antipodes Island, Macquarie Island, Auckland Islands and one sample from South Georgia. We find that most of the mice on the Kerguelen Archipelago have the same mitochondrial haplotype and all share the same major Y-chromosomal haplotype. Two small islands (Cochons Island and Cimetière Island) within the archipelago show a different mitochondrial haplotype, are genetically distinct for autosomal loci, but share the major Y-chromosomal haplotype. In the mitochondrial D-loop sequences, we find several single step mutational derivatives of one of the major mitochondrial haplotypes, suggesting an unusually high mutation rate, or the occurrence of selective sweeps in mitochondria. CONCLUSIONS: Although there was heavy ship traffic for over a hundred years to the Kerguelen Archipelago, it appears that the mice that have arrived first have colonized the main island (Grande Terre) and most of the associated small islands. The second invasion that we see in our data has occurred on islands that are detached from Grande Terre and were likely to have had no resident mice prior to their arrival. The genetic data suggest that the mice of both primary invasions originated from related source populations. Our data suggest that an area colonized by mice is refractory to further introgression, possibly due to fast adaptations of the resident mice to local conditions.