Bayesian analyses of admixture in wild and domestic cats (Felis silvestris) using linked microsatellite loci

Methods recently developed to infer population structure and admixture mostly use individual genotypes described by unlinked neutral markers. However, Hardy-Weinberg and linkage disequilibria among independent markers decline rapidly with admixture time, and the admixture signals could be lost in a few generations. In this study, we aimed to describe genetic admixture in 182 European wild and domestic cats (Felis silvestris), which hybridize sporadically in Italy and extensively in Hungary. Cats were genotyped at 27 microsatellites, including 21 linked loci mapping on five distinct feline linkage groups. Genotypes were analysed with structure 2.1, a Bayesian procedure designed to model admixture linkage disequilibrium, which promises to assess efficiently older admixture events using tightly linked markers. Results showed that domestic and wild cats sampled in Italy were split into two distinct clusters with average proportions of membership Q > 0.90, congruent with prior morphological identifications. In contrast, free-living cats sampled in Hungary were assigned partly to the domestic and the wild cat clusters, with Q < 0.50. Admixture analyses of individual genotypes identified, respectively, 5/61 (8%), and 16-20/65 (25-31%) hybrids among the Italian wildcats and Hungarian free-living cats. Similar results were obtained in the past using unlinked loci, although the new linked markers identified additional admixed wildcats in Italy. Linkage analyses confirm that hybridization is limited in Italian, but widespread in Hungarian wildcats, a population that is threatened by cross-breeding with free-ranging domestic cats. The total panel of 27 loci performed better than the linked loci alone in the identification of domestic and known hybrid cats, suggesting that a large number of linked plus unlinked markers can improve the results of admixture analyses. Inferred recombination events led to identify the population of origin of chromosomal segments, suggesting that admixture mapping experiments can be designed also in wild populations.     

Morphological discriminants of Scottish wildcats (Felis silvestris), domestic cats (F. catus) and their hybrids

Skull characters of wildcats ( Felis silvestris ), classed as 'old', 'recent' or 'modern', depending on collection date, were compared with those of domestic cats ( F. catus ) and hybrids, using Fisher Linear Discriminant Functions, Principal Component Analyses and Canonical Variates. A key is derived for classification of individual (unknown) skulls. All analyses indicate that 'recent' and 'modern' wildcats were different from those collected earlier, and implied marked changes in the rate of hydridization during this century. We suggest that the older wildcat populations were a relatively 'pure' form but more recent populations contain a significant hybrid component. Although there may be a trend towards re-establishment of the earlier type, an alternative conclusion is that the 'pure' form of wildcat is effectively extinct in Scotland.     

Taste reactivity patterns in domestic cats (Felis silvestris catus)

The present study is aimed at finding taste reactivity patterns in domestic cats (Felis silvestris catus) which reflect 'liking' or perceived palatability. Three groups of nonstressed cats living in households were formed which a priori were expected to differ in motivational state for eating food items (more or less hungry), and which were offered two different food items differing in general taste properties (more or less flavourful food, MFF and LFF, respectively) around the time that they were fed their normal food. Analysis of the amount of food eaten showed that MFF was consumed regardless of hunger level and that LFF was consumed depending on the hunger level of cats: the more hungry cats ate more of LFF than the less hungry cats. Analysis of post-meal behavioural sequences showed that the 'MFF consumption sequence' differed from the 'LFF refusal sequence' and that the 'LFF consumption sequence' strongly resembled the 'MFF consumption sequence' but also contained elements of the 'LFF refusal sequence'. Subsequent analysis of the frequencies and total durations of behavioural patterns showed that two kinds of patterns existed, possibly reflecting two 'palatability dimensions': hedonic taste reactivity patterns (lick/sniff feeding bowl, lip lick and groom face) and aversive taste reactivity patterns (lick/sniff food and lick nose). These dimensions may be combined to obtain a single palatability score.     

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.     

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.     

Clusters in social behaviour of female domestic cats (Felis silvestris catus) living in confinement

Associations between different agonistic and affiliative behavioural patterns of female domestic cats (Felis silvestris catus) were studied. In three groups of intact cats living in confinement frequencies of fourteen agonistic and affiliative behavioural patterns were recorded. The technique of factor analysis (Principal Components Analysis followed by varimax rotation on a dyads X behavioural patterns matrix) was used to detect clusters in these behavioural patterns. Five factors (or types of interindividual relationships) were extracted per group. They accounted collectively for at least 77% of the total variance present in the data. Although differences existed between groups with respect to behavioural patterns included in each factor, four clusters of behaviours could be discriminated: (I) social rubbing, lordosis and rolling in front of partner (sexual behaviour), (II) allogrooming, social sniffing, nosing, sniffing rear and treading (inspection-affiliative behaviour), (III) offensive behaviour and staring, and (IV) defensive behaviour and staring. The role of these clusters in group living is discussed.     

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.     

When domestic cat (Felis silvestris catus) population structures interact with their viruses

Many theoretical studies have proposed different causal mechanisms by which the structure of a host population could have important implications for life history traits of pathogens. However, little information is available from real systems to test these hypotheses. The domestic cat, Felis silvestris catus, whose populations exhibit a great variability in social and spatial structure, represent an ideal case study to assess this question. In the present article, we show how cat population structure may have influenced the evolution of feline viruses and, in return, how these viruses may have modified the genetic structure of cat populations. To cite this article: D. Pontier et al., C. R. Biologies 332 (2009).     

Differences in food preferences between individuals and populations of domestic cats Felis silvestris catus

The food preferences of 64 cats, of which 28 were domestic pets and 36 were free-ranging animals on three farms, were tested using five food types; a sixth type was also tested on the farm cats. Information was also gathered on the background diet of the house cats (individually) and of each farm cat colony. Consistent differences were found between the predefined groups of cats in their preferences for three foods, Hard Dry (HD), Canned Meat (CM) and Raw Beef (RB), the greatest differences being between house cats and farm cats as a whole. The three colonies of farm cats showed distinct differences in preference, for RB and/or CM, and for the sixth food, Soft Dry (SD). Differences between farms could be explained by a reduced preference for items similar to major components of the background diet of each colony, i.e. selection in favour of foods that were temporally rare. Differences between house and farm cats, and within the house cats, could not be explained in this way; the house cats were neophobic towards RB, and the farm cats ate little of the HD food, possibly because they found it difficult to ingest.     

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.     

Artificial insemination in domestic cats (Felis catus)

Artificial insemination (AI) in cats represents an important technique for increasing the contribution of genetically valuable individuals in specific populations, whether they be highly pedigreed purebred cats, medically important laboratory cats or endangered non-domestic cats. Semen is collected using electrical stimulation, with an artificial vagina or from intact or excised cauda epididymis. Sperm samples can be used for AI immediately after collection, after temporary storage above 0 degrees C or after cryopreservation. There have been three and five reports on intravaginal and intrauterine insemination, respectively, and one report on tubal insemination with fresh semen. In studies using fresh semen, it was reported that conception rates of 50% or higher were obtained by intravaginal insemination with 10-50x10(6) spermatozoa, while, in another report, the conception rate was 78% after AI with 80x10(6) spermatozoa. After intrauterine insemination, conception rates following deposition of 6.2x10(6) and 8x10(6) spermatozoa were reported to be 50 and 80%, respectively. With tubal insemination, the conception rate was 43% when 4x10(6) spermatozoa were used, showing that the number of spermatozoa required to obtain a satisfactory conception rate was similar to that of cats inseminated directly into the uterus. When frozen semen was used for intravaginal insemination the conception rate was rather low, but intrauterine insemination with 50x10(6) frozen/thawed spermatozoa resulted in a conception rate of 57%. Furthermore, in one report, conception was obtained by intrauterine insemination of frozen epididymal spermatozoa. Overall, there have been few reports on artificial insemination in cats. The results obtained to date show considerable variation, both within and among laboratories depending upon the type and number of spermatozoa used and the site of sperm deposition. Undoubtedly, future studies will identify the major factors required to consistently obtain reliable conception rates, so that AI can become a practical technique for enhancing the production of desirable genotypes, both for laboratory and conservation purposes.     

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.     

Paw preferences in cats (Felis silvestris catus) living in a household environment

Unrestrained, naı̈ve cats (Felis silvestris catus) (n=48: 28 males and 20 females), living in a natural domestic environment, were studied for paw preferences using a food reaching test. A total of 46% were right-preferent, 44% were left-preferent and 10% were ambilateral. 60% of the cats in our sample used one paw 100% of the time. This preference was stable over time (10 weeks), and was not influenced by the presence of food residue on the cats' non-preferred paw. There was no difference between male and female cats in the proportions of left and right paw-preferent individuals. We have reviewed the literature reporting paw preferences in cats and conclude that in static food-reaching tests, domestic cats show a marked paw preference with an equal distribution of left- and right-preferent individuals. They do not, however, show a group or population bias toward the use of any one paw. There are no significant sex differences. In contrast, there is previously published evidence which appears to suggest that moving-target reaching tests uncover a left-sided behavioural asymmetry.     

The function of allogrooming in domestic cats (Felis silvestris catus); a study in a group of cats living in confinement

Grooming interactions (n=83) occurring in a group of non free-ranging adult neutered male (n=14) and female (n=11) domestic cats (Felis silvestris catus) were analysed. Grooming was not induced by the proximity (distance <=0.5 m) of another animal. Grooming was in general directed at the head-neck area. Higher ranking animals groomed lower ranking animals more often than the other way round. Groomers tended to adopt ‘higher’ (standing, sitting upright) postures than groomees (sitting, lying). Agonistic behaviour occurred in 35% of interactions. Groomers showed offensive behaviour more often than groomees, most often after grooming a partner. Furthermore groomers often groomed themselves after grooming a partner. The degree of relatedness of animals did not affect the frequencies or durations of grooming. These results are consistent with the hypothesis that allogrooming in domestic cats may be a way of redirecting (potential) aggression in situations in which overt aggression is too costly. The data were previously presented at the 29th International Congress of the International Society for Applied Ethology (van den Bos, R. (1995) Allogrooming in domestic cats in confinement, Proceedings of the 29th International Congress of the International Society for Applied Ethology, S.M. Rutter et al. (Eds.), pp. 109–110)     

Naturally occurring Sarcocystis infection in domestic cats (Felis catus)

Equine protozoal myeloencephalitis is an important neurological disease of horses in the United States. Consequently, there is an active research effort to identify hosts associated with the primary causative agent, Sarcocystis neurona. The purpose of this study was to determine whether the domestic cat (Felis catus) is a natural host for S. neurona. Muscle sections from 50 primarily free-roaming domestic cats were examined for the presence of sarcocysts. Serum from cats in this group and another group of 50 free-roaming cats were evaluated for the presence of S. neurona antibody. Sarcocysts were found in five of 50 (10%) cats, and S. neurona antibody in five of 100 (5%) cats. Morphological, molecular (including ribosomal RNA genes), and biological characterisation of these sarcocysts showed that they were not S. neurona or S. neurona-like. Sarcocysts found in the cats were identified morphologically as Sarcocystis felis, a common parasite of wild felids. The life cycle of S. felis is not known, and prior to this study, no molecular marker for S. felis existed. Although cats were found to be infected with S. felis sarcocysts, serological data provided evidence of possible infection with S. neurona as well. Further work is needed to determine the role of the domestic cat in the life cycle of S. neurona.     

The social function of tail up in the domestic cat (Felis silvestris catus)

We investigated the social function of tail up in order to verify its possible relationship with the hierarchical organization of a social group. Domestic cats live at higher densities than their ancestor which is a solitary species. Since the signals needed by solitary animals have different properties than those needed by group-living individuals, signalling pattern utilised by the domestic cat has inevitably changed. Kittens displayed the tail up when greeting their mother; this behaviour can also be observed in wild species. But, in domestic cat the tail up can be also observed when an adult individual meets another one and it signals the intention to interact amicably. Rank order affected the display of tail up posture: it was more frequently displayed by low-ranking cats, and high-ranking individuals received it more often than other members of the social group. Then, tail up seems to be a signal by means of which a cat shows the recognition of the higher social status of the individual to whom is directed. We confirmed the association between tail up and other affiliative behavioural patterns and the individual variability in displaying them. Considerations on the evolution of the tail up as a visual signal will be discussed.     

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.