Genetic identification of wild and domestic cats (Felis silvestris) and their hybrids using Bayesian clustering methods

Crossbreeding with free-ranging domestic cats is supposed to threaten the genetic integrity of wildcat populations in Europe, although the diagnostic markers to identify "pure" or "admixed" wildcats have never been clearly defined. Here we use mitochondrial (mt) DNA sequences and allelic variation at 12 microsatellite loci to genotype 128 wild and domestic cats sampled in Italy which were preclassified into three separate groups: European wildcats (Felis silvestris silvestris), Sardinian wildcats (Felis silvestris libyca), and domestic cats (Felis silvestris catus), according to their coat color patterns, collection localities, and other phenotypical traits, independently of any genetic information. For comparison, we included some captive-reared hybrids of European wild and domestic cats. Genetic variability was significantly partitioned among the three groups (mtDNA estimate of F(ST) = 0.36; microsatellite estimate of R(ST) = 0.30; P < 0.001), suggesting that morphological diversity reflects the existence of distinct gene pools. Multivariate ordination of individual genotypes and clustering of interindividual genetic distances also showed evidence of distinct cat groups, partially congruent with the morphological classification. Cluster analysis, however, did not enable hybrid cats to be identified from genetic information alone, nor were all individuals assigned to their populations. In contrast, a Bayesian admixture analysis simultaneously assigned the European wildcats, the Sardinian wildcats, and the domestic cats to different clusters, independent of any prior information, and pointed out the admixed gene composition of the hybrids, which were assigned to more than one cluster. Only one putative Sardinian wildcat was assigned to the domestic cat cluster, and one presumed European wildcat showed mixed (hybrid) ancestry in the domestic cat gene pool. Mitochondrial DNA sequences indicated that three additional presumed European wildcats might have hybrid ancestry. These four cats were sampled from the same area in the northernmost edge of the European wildcat distribution in the Italian Apennines. Admixture analyses suggest that wild and domestic cats in Italy are distinct, reproductively isolated gene pools and that introgression of domestic alleles into the wild-living population is very limited and geographically localized.     

Genetic analysis shows low levels of hybridization between African wildcats (Felis silvestris lybica) and domestic cats (F. s. catus) in South Africa

Hybridization between domestic and wild animals is a major concern for biodiversity conservation, and as habitats become increasingly fragmented, conserving biodiversity at all levels, including genetic, becomes increasingly important. Except for tropical forests and true deserts, African wildcats occur across the African continent; however, almost no work has been carried out to assess its genetic status and extent of hybridization with domestic cats. For example, in South Africa it has been argued that the long‐term viability of maintaining pure wildcat populations lies in large protected areas only, isolated from human populations. Two of the largest protected areas in Africa, the Kgalagadi Transfrontier and Kruger National Parks, as well as the size of South Africa and range of landscape uses, provide a model situation to assess how habitat fragmentation and heterogeneity influences the genetic purity of African wildcats. Using population genetic and home range data, we examined the genetic purity of African wildcats and their suspected hybrids across South Africa, including areas within and outside of protected areas. Overall, we found African wildcat populations to be genetically relatively pure, but instances of hybridization and a significant relationship between the genetic distinctiveness (purity) of wildcats and human population pressure were evident. The genetically purest African wildcats were found in the Kgalagadi Transfrontier Park, while samples from around Kruger National Park showed cause for concern, especially combined with the substantial human population density along the park's boundary. While African wildcat populations in South Africa generally appear to be genetically pure, with low levels of hybridization, our genetic data do suggest that protected areas may play an important role in maintaining genetic purity by reducing the likelihood of contact with domestic cats. We suggest that approaches such as corridors between protected areas are unlikely to remain effective for wildcat conservation, as the proximity to human settlements around these areas is projected to increase the wild/domestic animal interface. Thus, large, isolated protected areas will become increasingly important for wildcat conservation and efforts need to be made to prevent introduction of domestic cats into these areas.     

Genetic distinction of wildcat (Felis silvestris) populations in Europe,and hybridization with domestic cats in Hungary

The genetic integrity and evolutionary persistence of declining wildcat populations are threatened by crossbreeding with widespread free-living domestic cats. Here we use allelic variation at 12 microsatellite loci to describe genetic variation in 336 cats sampled from nine European countries. Cats were identified as European wildcats (Felis silvestris silvestris), Sardinian wildcats (F. s. libyca) and domestic cats (F. s. catus), according to phenotypic traits, geographical locations and independently of any genetic information. Genetic variability was significantly partitioned among taxonomic groups (FST = 0.11; RST = 0.41; P < 0.001) and sampling locations (FST = 0.07; RST = 0.06; P < 0.001), suggesting that wild and domestic cats are subdivided into distinct gene pools in Europe. Multivariate and Bayesian clustering of individual genotypes also showed evidence of distinct cat groups, congruent with current taxonomy, and suggesting geographical population structuring. Admixture analyses identified cryptic hybrids among wildcats in Portugal, Italy and Bulgaria, and evidenced instances of extensive hybridization between wild and domestic cats sampled in Hungary. Cats in Hungary include a composite assemblage of variable phenotypes and genotypes, which, as previously documented in Scotland, might originate from long lasting hybridization and introgression. A number of historical, demographic and ecological conditions can lead to extensive crossbreeding between wild and domestic cats, thus threatening the genetic integrity of wildcat populations in Europe.     

Low rates of hybridization between European wildcats and domestic cats in a human‐dominated landscape

Hybridization between wild species and their domestic congeners is considered a major threat for wildlife conservation. Genetic integrity of the European wildcat, for instance, is a concern as they are outnumbered by domestic cats by several orders of magnitude throughout its range. We genotyped 1,071 individual wildcat samples obtained from hair traps and roadkills collected across the highly fragmented forests of western Central Europe, in Germany and Luxembourg, to assess domestic cat introgression in wildcats in human‐dominated landscapes. Analyses using a panel of 75 autosomal SNPs suggested a low hybridization rate, with 3.5% of wildcat individuals being categorized as F1, F2, or backcrosses to either parental taxon. We report that results based on a set of SNPs were more consistent than on a set of 14 microsatellite markers, showed higher accuracy to detect hybrids and their class in simulation analyses, and were less affected by underlying population structure. Our results strongly suggest that very high hybridization rates previously reported for Central Europe may be partly due to inadequate choice of markers and/or sampling design. Our study documents that an adequately selected SNP panel for hybrid detection may be used as an alternative to commonly applied microsatellite markers, including studies relying on noninvasively collected samples. In addition, our finding of overall low hybridization rates in Central European wildcats provides an example of successful wildlife coexistence in human‐dominated, fragmented landscapes.     

European wildcat populations are subdivided into five main biogeographic groups: consequences of Pleistocene climate changes or recent anthropogenic fragmentation?

Extant populations of the European wildcat are fragmented across the continent, the likely consequence of recent extirpations due to habitat loss and over‐hunting. However, their underlying phylogeographic history has never been reconstructed. For testing the hypothesis that the European wildcat survived the Ice Age fragmented in Mediterranean refuges, we assayed the genetic variation at 31 microsatellites in 668 presumptive European wildcats sampled in 15 European countries. Moreover, to evaluate the extent of subspecies/population divergence and identify eventual wild × domestic cat hybrids, we genotyped 26 African wildcats from Sardinia and North Africa and 294 random‐bred domestic cats. Results of multivariate analyses and Bayesian clustering confirmed that the European wild and the domestic cats (plus the African wildcats) belong to two well‐differentiated clusters (average Ф_ST = 0.159, R_st  = 0.392, P > 0.001; Analysis of molecular variance [AMOVA]). We identified from c. 5% to 10% cryptic hybrids in southern and central European populations. In contrast, wild‐living cats in Hungary and Scotland showed deep signatures of genetic admixture and introgression with domestic cats. The European wildcats are subdivided into five main genetic clusters (average Ф_ST = 0.103, R_st  = 0.143, P > 0.001; AMOVA) corresponding to five biogeographic groups, respectively, distributed in the Iberian Peninsula, central Europe, central Germany, Italian Peninsula and the island of Sicily, and in north‐eastern Italy and northern Balkan regions (Dinaric Alps). Approximate Bayesian Computation simulations supported late Pleistocene–early Holocene population splittings (from c. 60 k to 10 k years ago), contemporary to the last Ice Age climatic changes. These results provide evidences for wildcat Mediterranean refuges in southwestern Europe, but the evolution history of eastern wildcat populations remains to be clarified. Historical genetic subdivisions suggest conservation strategies aimed at enhancing gene flow through the restoration of ecological corridors within each biogeographic units. Concomitantly, the risk of hybridization with free‐ranging domestic cats along corridor edges should be carefully monitored.     

Genetic structure of wildcat (Felis silvestris) populations in Italy

Severe climatic changes during the Pleistocene shaped the distributions of temperate‐adapted species. These species survived glaciations in classical southern refuges with more temperate climates, as well as in western and eastern peripheral Alpine temperate areas. We hypothesized that the European wildcat (Felis silvestris silvestris) populations currently distributed in Italy differentiated in, and expanded from two distinct glacial refuges, located in the southern Apennines and at the periphery of the eastern Alps. This hypothesis was tested by genotyping 235 presumed European wildcats using a panel of 35 domestic cat‐derived microsatellites. To provide support and controls for the analyses, 17 know wildcat x domestic cat hybrids and 17 Sardinian wildcats (F. s. libyca) were included. Results of Bayesian clustering and landscape genetic analyses showed that European wildcats in Italy are genetically subdivided into three well‐defined clusters corresponding to populations sampled in: (1) the eastern Alps, (2) the peninsular Apennines, and (3) the island of Sicily. Furthermore, the peninsular cluster is split into two subpopulations distributed on the eastern (Apennine mountains and hills) and western (Maremma hills and lowlands) sides of the Apennine ridge. Simulations indicated Alpine, peninsular, and Sicilian wildcats were isolated during the Last Glacial Maximum. Population subdivision in the peninsula cluster of central Italy arose as consequence of a more recent expansions of historically or ecologically distinct European wildcat subpopulations associated with distinct the Continental or Mediterranean habitats. This study identifies previously unknown European wildcat conservation units and supports a deep phylogeographical history for Italian wildcats.     

Molecular analysis of hybridisation between wild and domestic cats (Felis silvestris) in Portugal: implications for conservation

The endangered European wildcat (Felis silvestris silvestris) is represented, today, by fragmented and declining populations whose genetic integrity is considered to be seriously threatened by crossbreeding with widespread free-ranging domestic cats. Extensive and recent hybridisation has been described in Hungary and Scotland, in contrast with rare introgression of domestic alleles in Italy and Germany. In Portugal, the wildcat is now listed as VULNERABLE in the Red Book of Portuguese Vertebrates. Nevertheless, genetic diversity of populations and the eventual interbreeding with domestic cats remain poorly studied. We surveyed genetic variation at 12 autosomal microsatellites for 34 wild and 64 domestic cats collected across Portugal. Wild and domestic cats were significantly differentiated both at allele frequencies and sizes (F _ST=0.11, R _ST = 0.18, P < 0.001). Population structure and admixture analyses performed using Bayesian approaches also showed evidence of two discrete groups clustering wild and domestic populations. Results did not show significant genetic divergence among Northern, Central and Southern wildcats. Six morphologically identified wildcats were significantly assigned to the domestic cluster, revealing some discrepancy between phenotypic and genetic identifications. We detected four hybrids (approximately 14%) using a consensus analysis of different Bayesian model-based software. These hybrids were identified throughout all sampled areas, suggesting that hybridisation is of major concern for the appropriate implementation of wildcat conservation strategies in Portugal.     

Hybridization versus conservation: are domestic cats threatening the genetic integrity of wildcats (Felis silvestris silvestris) in Iberian Peninsula?

Cross-breeding between wild and free-ranging domestic species is one of the main conservation problems for some threatened species. The situation of wildcats (Felis silvestris silvestris) in Europe is a good example of this critical phenomenon. Extensive hybridization was described in Hungary and Scotland, contrasting with occasional interbreeding in Italy and Germany. First analyses in Portugal revealed a clear genetic differentiation between wild and domestic cats; however, four hybrids were detected. Here, we extended the approach to Iberian Peninsula using multivariate and Bayesian analyses of multilocus genotypes for 44 Portuguese wildcats, 31 Spanish wildcats and 109 domestic cats. Globally, wild and domestic cats were significantly differentiated (FST=0.20, p<0.001) and clustered into two discrete groups. Diverse clustering methods and assignment criteria identified an additional hybrid in Portugal, performing a total of five admixed individuals. The power of admixture analyses was assessed by simulating hybrid genotypes, which revealed that used microsatellites were able to detect 100, 91 and 85% of first-generation hybrids, second-generation genotypes and backcrosses, respectively. These findings suggest that the true proportion of admixture can be higher than the value estimated in this study and that the improvement of genetic tools for hybrids detection is crucial for wildcat conservation.     

Regionally high rates of hybridization and introgression in German wildcat populations (Felis silvestris, Carnivora, Felidae)

While the western populations of the wildcat ( Felis silvestris silvestris ) in Germany come into contact with wildcats in France and Switzerland, the eastern distribution area is geographically completely isolated and consists of scattered subpopulations. To investigate population structure, evolutionary relationships and degree of hybridization with domestic cats we analysed the mitochondrial control region of 86 cats in combination with 11 microsatellite loci of 149 cats. According to our microsatellite data, German wildcats are divided into two separate populations corresponding to the western and eastern distribution areas. We found no indication of a further subdivision of the eastern population. German wildcat populations are genetically distinct from domestic cats in the main, but we identified 18.4% of the whole wildcat sample as being of hybrid origin, corresponding to 4.2% of the eastern and 42.9% of the western wildcat population, and 2.7% of the domestic cat sample. The mitochondrial haplotypes form a network of three connected clusters and reveal a high level of genetic diversity, especially within the eastern population. Our findings are explained at best in terms of continuous introgression between domestic cats and wildcat populations and differing degrees of recent hybridization in the various populations. Future conservation efforts should focus on preserving the existing gene flow between the isolated distribution areas, but also on preventing the spread of hybrids and limiting the habitat alterations that lead to increased contact with domestic cats. In conclusion we discuss possible evolutionary reasons for the still traceable genetic integrity of the wildcat despite its long history of interbreeding.     

Strong spatial segregation between wildcats and domestic cats may explain low hybridization rates on the Iberian Peninsula

The European wildcat (Felis silvestris silvestris) is an endangered felid impacted by genetic introgression with the domestic cat (Felis silvestris catus). The problem of hybridization has had different effects in different areas. In non-Mediterranean regions pure forms of wildcats became almost extinct, while in Mediterranean regions genetic introgression is a rare phenomenon. The study of the potential factors that prevent the gene flow in areas of lower hybridization may be key to wildcat conservation. We studied the population size and spatial segregation of wildcats and domestic cats in a typical Mediterranean area of ancient sympatry, where no evidence of hybridization had been detected by genetic studies. Camera trapping of wild-living cats and walking surveys of stray cats in villages were used for capture–recapture estimations of abundance and spatial segregation. Results showed (i) a low density of wildcats and no apparent presence of putative hybrids; (ii) a very low abundance of feral cats in spite of the widespread and large population sources of domestic cats inhabiting villages; (iii) strong spatial segregation between wildcats and domestic/feral cats; and (iv) no relationship between the size of the potential population sources and the abundance of feral cats. Hence, domestic cats were limited in their ability to become integrated into the local habitat of wildcats. Ecological barriers (habitat preferences, food limitations, intra-specific and intra-guild competition, predation) may explain the severe divergences of hybridization impact observed at a biogeographic level. This has a direct effect on key conservation strategies for wildcats (i.e., control of domestic cats).     

Female in the inside,male in the outside: insights into the spatial organization of a European wildcat population

Hybridization between the European wildcat, Felis silvestris silvestris, and the domestic cat, Felis silvestris catus, has been found in several European countries with different landscape structures and in various proportions. In this study, we focus on a local population of European wildcats in forests fragmented by agricultural lands in northeastern France. Our aim is to better understand how the spatial organization of the wildcats in this particular type of environment might impact the proportion of hybridization. We combined radio-tracking and genetics through the use of microsatellite markers in order to assess both the spacing pattern and the level of hybridization of this wildcat population. Hybridization is rare in this wildcat population with only one putative hybrid (most likely backcrossed) detected out of 42 putative wildcats. We found that most females were concentrated inside the forest while males stood in the periphery or outside the forest. Furthermore, many males and females resulted related. Such a spacing pattern might limit contacts between male domestic cats and female wildcats and can be one of the causes that explain the low level of hybridization in the wildcat population in this environment. We could not exclude the possibility of hybrid presence in the neighboring domestic cat populations. Our results yield new insights on the influence that the landscape configuration and the spacing pattern can have on genetic flow between the populations of the two subspecies.     

Craniological differentiation between European wildcats (Felis silvestris silvestris), African wildcats (F. s. lybica) and Asian wildcats (F. s. ornata): implications for their evolution and conservation

Intraspecific diversification of the wildcat (Felis silvestris), including the European wildcat (F. s. silvestris), the Asian wildcat (F. s. ornata) and the African wildcat (F. s. lybica), was examined based on 39 cranial morphology variables. The samples of free‐ranging cats originated from Britain, Europe, Central Asia and southern Africa, consisting of both nominal wildcat specimens (referred to henceforth as ‘wildcats’) and nominal non‐wildcat specimens (‘non‐wildcats’) based on museum labels. The skull morphology of ‘wildcats’ from Britain and Europe is clearly different from that of ‘wildcats’ of Central Asia and southern Africa. The latter are characterized especially by their proportionately larger cheek teeth. On the basis of principal component, discriminant function and canonical variate analyses, the skull morphology of British ‘non‐wildcats’ is less distinct than is that of British ‘wildcats’ from the skull morphologies of ‘wildcats’ of Central Asia and southern Africa. On the other hand, the skull morphology of southern African ‘non‐wildcats’ is as distinct from those of ‘wildcats’ of Britain and Europe as is that of southern African ‘wildcats’. We suggest that the evolution of the modern wildcat probably consisted of at least three different distribution expansions punctuated by two differentiation events: the exodus from Europe during the late Pleistocene, coinciding with the emergence of the steppe wildcat lineage (phenotype of Asian–African wildcat), followed by its rapid range expansion in the Old World. The second differentiation event was the emergence of the domestic cat followed by its subsequent colonization of the entire world with human assistance. Considering the recent evolutionary history of, and morphological divergence in, the wildcat, preventing hybridization between the European wildcat and the domestic cat is a high conservation priority. © 2004 The Linnean Society of London, Biological Journal of the Linnean Society, 2004, 83 , 47–63.     

Wildcat population density in NE Portugal: A regional stronghold for a nationally threatened felid

Population density data on depleted and endangered wildlife species are essential to assure their effective management and, ultimately, conservation. The European wildcat is an elusive and threatened species inhabiting the Iberian Peninsula, with fragmented populations and living in low densities. We fitted spatial capture–recapture models on camera-trap data, to provide the first estimate of wildcat density for Portugal and assess the most influential drivers determining it. The study was implemented in Montesinho Natural Park (NE Portugal), where we identified nine individuals, over a total effort of 3,477 trap-nights. The mean density estimate was 0.032 ± 0.012 wildcat/km², and density tended to increase with distance to humanized areas, often linked to lower human disturbance and domestic cat presence, with forest and herbaceous vegetation cover and with European rabbit abundance. Although, this density estimate is within the range of values estimated for protected areas elsewhere in the Iberian Peninsula, our estimates are low at the European level. When put in context, our results highlight that European wildcats may be living in low population densities across the Iberian Mediterranean biogeographic region. No phenotypic domestic or hybrid cats were detected, suggesting potentially low admixture rates between the two species, although genetic sampling would be required to corroborate this assertion. We provide evidence that Montesinho Natural Park may be a suitable area to host a healthy wildcat population, and thus be an important protected area in this species' conservation context.     

Toward a genome-wide approach for detecting hybrids: informative SNPs to detect introgression between domestic cats and European wildcats (Felis silvestris)

Endemic gene pools have been severely endangered by human-mediated hybridization, which is posing new challenges in the conservation of several vertebrate species. The endangered European wildcat is an example of this problem, as several natural populations are suffering introgression of genes from the domestic cat. The implementation of molecular methods for detecting hybridization is crucial for supporting appropriate conservation programs on the wildcat. In this study, genetic variation at 158 single-nucleotide polymorphisms (SNPs) was analyzed in 139 domestic cats, 130 putative European wildcats and 5 captive-bred hybrids (N=274). These SNPs were variable both in wild (H_E=0.107) and domestic cats (H_E=0.340). Although we did not find any SNP that was private in any population, 22 SNPs were monomorphic in wildcats and pairwise F_CT values revealed marked differences between domestic and wildcats, with the most divergent 35 loci providing an average F_CT>0.74. The power of all the loci to accurately identify admixture events and discriminate the different hybrid categories was evaluated. Results from simulated and real genotypes show that the 158 SNPs provide successful estimates of admixture, with 100% hybrid individuals (two to three generations in the past) being correctly identified in STRUCTURE and over 92% using the NEWHYBRIDS'' algorithm. None of the unclassified cats were wrongly allocated to another hybrid class. Thirty-five SNPs, showing the highest F_CT values, provided the most parsimonious panel for robust inferences of parental and first generations of admixed ancestries. This approach may be used to further reconstruct the evolution of wildcat populations and, hopefully, to develop sound conservation guidelines for its legal protection in Europe.     

Development of SNP markers identifying European wildcats,domestic cats,and their admixed progeny

Introgression can be an important evolutionary force but it can also lead to species extinction and as such is a crucial issue for species conservation. However, introgression is difficult to detect, morphologically as well as genetically. Hybridization with domestic cats (Felis silvestris catus) is a major concern for the conservation of European wildcats (Felis s. silvestris). The available morphologic and genetic markers for the two Felis subspecies are not sufficient to reliably detect hybrids beyond first generation. Here we present a single nucleotide polymorphism (SNP) based approach that allows the identification of introgressed individuals. Using high‐throughput sequencing of reduced representation libraries we developed a diagnostic marker set containing 48 SNPs (Fst  > 0.8) which allows the identification of wildcats, domestic cats, their hybrids and backcrosses. This allows assessing introgression rate in natural wildcat populations and is key for a better understanding of hybridization processes.     

Spatial colonization by feral domestic catsFelis catus of former wildcatFelis silvestris silvestris home ranges

Presently, wildcatFelis silvestris silvestris Schreber, 1777 populations are fragmented and rapidly declining in most of Europe. Although habitat destruction possibly constitutes the most serious threat to wildcat survival, hybridisation with feral domestic cats is also a critical problem. However, the mechanisms that allow domestic cats to colonise former wild cat home ranges are yet unclear. The present paper describes the decrease of typical phenotypic wildcats and the increase of phenotypic domestic cats in a remote wild area of Portugal (Serra da Malcata). A field survey using box-traps and radio-tracking between 1998 and 2001 revealed that wildcats were widespread in the study area and no domestic cats were present. A second survey using camera traps between 2005 and 2007 revealed only one wildcat whereas four typical domestic phenotype individuals were photographed. The present study clearly emphasizes the need for urgent measures aimed at preserving wildcat populations. These measures should include a national census of the species and an extensive monitoring of genetic integrity of wildcat populations, followed by the elaboration of a wildcat conservation action plan.     

The Genetic Integrity of the Ex Situ Population of the European Wildcat (Felis silvestris silvestris) Is Seriously Threatened by Introgression from Domestic Cats (Felis silvestris catus)

Studies on the genetic diversity and relatedness of zoo populations are crucial for implementing successful breeding programmes. The European wildcat, Felis s. silvestris, is subject to intensive conservation measures, including captive breeding and reintroduction. We here present the first systematic genetic analysis of the captive population of Felis s. silvestris in comparison with a natural wild population. We used microsatellites and mtDNA sequencing to assess genetic diversity, structure and integrity of the ex situ population. Our results show that the ex situ population of the European wildcat is highly structured and that it has a higher genetic diversity than the studied wild population. Some genetic clusters matched the breeding lines of certain zoos or groups of zoos that often exchanged individuals. Two mitochondrial haplotype groups were detected in the in situ populations, one of which was closely related to the most common haplotype found in domestic cats, suggesting past introgression in the wild. Although native haplotypes were also found in the captive population, the majority (68%) of captive individuals shared a common mtDNA haplotype with the domestic cat (Felis s. catus). Only six captive individuals (7.7%) were assigned as wildcats in the STRUCTURE analysis (at K = 2), two of which had domestic cat mtDNA haplotypes and only two captive individuals were assigned as purebred wildcats by NewHybrids. These results suggest that the high genetic diversity of the captive population has been caused by admixture with domestic cats. Therefore, the captive population cannot be recommended for further breeding and reintroduction.     

Genetic diversity and integrity of German wildcat (Felis silvestris) populations as revealed by microsatellites,allozymes, and mitochondrial DNA sequences

As a consequence of persecution and habitat fragmentation, wildcats (Felis silvestris silvestris) in Western Europe have experienced a severe reduction in population numbers and sizes. The remaining wildcat populations are considered to be endangered by losses of genetic variability and by hybridisation with free-ranging domestic cats. To investigate genetic diversity within and among wild and domestic cat populations in Germany and to estimate the extent of gene flow between both forms, we analysed a total of 266 individuals. PCR-amplification and sequencing of 322 base pairs of a highly variable part of the mitochondrial control region (HV1) of 244 specimens resulted in 41 haplotypes with 31 polymorphic sites. Additionally, eight microsatellite loci were examined for those 244 cats. Moreover, a total of 46 wildcats and 22 domestic cats could be genotyped for 13 polymorphic out of 31 enzyme loci. Genetic variability in both groups was generally high. Variability in domestic cat populations was higher than in wildcat populations. Almost no differentiation between domestic cat populations could be found (F_ST for microsatellites=3%). In contrast, wildcat populations differed significantly from one another (F_ST for microsatellites=9.55%) Within the smaller wildcat populations, a reduction of genetic diversity was detectable with regard to the nuclear DNA. Wildcat and domestic cat mitochondrial haplotypes were separated, suggesting a very low level of maternal gene flow between both forms. In microsatellites and to a somewhat lesser extent in allozymes, wildcats and domestic cats showed distinct differentiation, suggesting an only low extent of past hybridisation in certain populations. The microsatellite data set indicated a significantly reduced effective population size (bottleneck) in the recent past for one German wildcat population.     

Evaluation of anatomical characters and the question of hybridization with domestic cats in the wildcat population of Thuringia, Germany

Germany's large population of wildcats ( Felis silvestris silvestris ) can be clearly distinguished from domestic cats on the basis of morphological characters. However, an examination of 71 specimens from Thuringia also illustrates the risks involved in using only a few such characters. The most reliable tool for identification in the field are three pelage characters (distinctness of tail bands, stripes on the nape and stripes on the shoulder). Only two morphological characters (intestine length and cranial volume) are unambiguous and demonstrate no overlap in distribution between domestic cats and wildcats. A linear discriminant analysis with forward selection of variables showed that only five skull variables are necessary to distinguish all four groups (subspecies × sex). Additionally, the high degree of correlation between most of the 49 variables examined (as indicated by Pearson's r correlation matrix) speaks against the utility of measuring such high numbers of characters in the future. Principal component analysis (PCA) enabled the subspecies to be separated clearly. The first PCA axis was highly correlated with variables characterizing overall body size, thus separating male and female into wildcats and domestic cats. Even when the chief differentiating characters are missing, the PCA still resulted in a good separation of subspecies. None of the genetically determined hybrids could have been deciphered unambiguously using the morphological characters still intact after a road death. Hybridization seems to occur whenever wildcats change their ecological function and become field cats. The impulse to hybridize seems to come much more from the wildcat side than the side of feral cats, and deforestation represents the major threat to the wildcat.     

Spatial organisation of European wildcats (Felis silvestris silvestris) in an agriculturally dominated landscape in Central Europe

For the first time, a telemetry study of European wildcats was conducted outside heavily forested habitats, in an agriculturally-dominated landscape, which is assumed to be a suboptimal habitat in Central Europe. According to the current knowledge of morphological and genetic diagnoses all captured wildcats were purebred, without any hint of recent interbreeding. Our results confirmed the regular use of open landscape habitats also by resident individuals. Sex and age ratios of wildcats, using the open landscapes, were balanced, and one successful reproduction event was recorded. Female annual home-ranges were smaller compared to those recorded in forested habitats. Male annual home-ranges were similar in size, whereas the core areas were smaller than those recorded in adjacent forested habitats. Males had the largest home-ranges during spring and the smallest in summer, whereas females had the largest home-ranges during summer and the smallest during winter/spring. The structure of the apparently purebred population with resident individuals and a reproduction event in an open cultivated landscape may be related to the close proximity (3.5–5 km) of a wildcat core distribution area. As a consequence of our results, the potential range of the European wildcat is increased, because it may occur in agricultural landscapes where sufficient shelter is available. This should be considered in environmental impact analyses and species monitoring.