An STR Forensic Typing System for Genetic Individualization of Domestic Cat ( Felis catus ) Samples

NCI-Frederick, Фредерик, Maryland, United States
Journal of Forensic Sciences (Impact Factor: 1.16). 10/2005; 50(5):1061-70. DOI: 10.1520/JFS2004317
Source: PubMed


A forensic genotyping panel of 11 tetranucleotide STR loci from the domestic cat was characterized and evaluated for genetic individualization of cat tissues. We first examined 49 candidate STR loci and their frequency assessment in domestic cat populations. The STR loci (3-4 base pair repeat motifs), mapped in the cat genome relative to 579 coding loci and 255 STR loci, are well distributed across the 18 feline autosomes. All loci exhibit Mendelian inheritance in a multi-generation pedigree. Eleven loci that were unlinked and were highly heterozygous in cat breeds were selected for a forensic panel. Heterozygosity values obtained for the independent loci, ranged from 0.60-0.82, while the average cat breed heterozygosity obtained for the 11 locus panel was 0.71 (range of 0.57-0.83). A small sample set of outbred domestic cats displayed a heterozygosity of 0.86 for the 11 locus panel. The power of discrimination of the panel is moderate to high in the cat breeds examined, with an average P(m) of 3.7E-06. The panel shows good potential for genetic individualization within outbred domestic cats with a P(m) of 5.31E-08. A multiplex protocol, designed for the co-amplification of the 11 loci and a gender-identifying locus, is species specific and robust, generating a product profile with as little as 0.125 nanograms of genomic DNA.

Download full-text


Available from: Victor A David,
  • Source
    • "We genotyped all samples with a 12-microsatellite loci in a single multiplex reaction (MeowPlex). This included as a standard component a sex-identifying sequence tagged site from the domestic cat Y-chromosome SRY gene, which was as part of the multiplex set not separable for this study (Butler et al. 2002, Menotti-Raymond et al. 2005, Menotti- Raymond et al. 2012). We sequenced a stretch (1,800 bp) of mitochondrial DNA genome corresponding to the ND5 and ND6 region using primers and a polymerase chain reaction (PCR) protocol developed by S. Hendrickson-Lambert (personal communication; Supporting Information 1). "
    [Show abstract] [Hide abstract]
    ABSTRACT: Invasive predators have a major impact on endemic island species; therefore, information about invasion dynamics are essential for implementing successful control measures. The introduction of feral cats onto Dirk Hartog Island, Western Australia, has had devastating effects, with presumably 10 of 13 native terrestrial mammal species being lost because of predation. Detailed records of historical introduction events were lacking; therefore, we analyzed genetic variation of the current population to gain information about past invasion dynamics and current gene-flow patterns. We analyzed the genetic structure and diversity of feral cats on the island and 2 mainland populations (Peron Peninsula and Steep Point). Analysis of mitochondrial DNA (ND5 and ND6) showed 2 primary haplotypes that we attribute to 2 main introduction events. Pairwise values indicated high connectivity on the island but some isolation to the mainland populations. Mitochondrial and nuclear data showed no evidence for genetic differentiation of island and mainland populations; however, kinship analyses rejected evidence for on-going immigration of members of the current cat populations. Overall, our data suggested that gene flow following the main introduction events ceased some years ago. Because current island populations appear to be reproductively isolated from mainland populations, a sufficiently large-scale eradication measure might successfully diminish feral cat populations long-term. © 2014 The Wildlife Society.
    Journal of Wildlife Management 08/2014; 78(6). DOI:10.1002/jwmg.739 · 1.73 Impact Factor
  • Source
    • "Chr. Asn., chromosomal assignment of locus in species of origin; NI, no information; T, tetranucleotide repeat; D, dinucleotide repeat; bp, base pairs; Na, number of alleles; Ne, number of effective alleles; HO, observed heterozygosity; HE, PIC, polymorphic information content; expected heterozygosity; PID (locus), probability of identity between unrelated individuals; PID Sibs (locus), probability of identity between siblings; Height ratio 1, first stutter peak/main allele peak; Height ratio 2, minus A peak/main allele peak; Height ratio 3, plus A peak/main allele peak; Height ratio 4, heterozygote allele peak/main allele peak; PIC, polymorphic information content; FIS, inbreeding coefficients. 1Williamson et al. (2002); 2Menotti-Raymond et al. (1999 2005); 3Singh et al. (2002); 4Janecka et al. (2008); *significance of Hardy-Weinberg test (*P < 0.05). "
    [Show abstract] [Hide abstract]
    ABSTRACT: In India, six landscapes and source populations that are important for long-term conservation of Bengal tigers (Panthera tigris tigris) have been identified. Except for a few studies, nothing is known regarding the genetic structure and extent of gene flow among most of the tiger populations across India as the majority of them are small, fragmented and isolated. Thus, individual-based relationships are required to understand the species ecology and biology for planning effective conservation and genetics-based individual identification has been widely used. But this needs screening and describing characteristics of microsatellite loci from DNA from good-quality sources so that the required number of loci can be selected and the genotyping error rate minimized. In the studies so far conducted on the Bengal tiger, a very small number of loci (n = 35) have been tested with high-quality source of DNA, and information on locus-specific characteristics is lacking. The use of such characteristics has been strongly recommended in the literature to minimize the error rate and by the International Society for Forensic Genetics (ISFG) for forensic purposes. Therefore, we describe for the first time locus-specific genetic and genotyping profile characteristics, crucial for population genetic studies, using high-quality source of DNA of the Bengal tiger. We screened 39 heterologous microsatellite loci (Sumatran tiger, domestic cat, Asiatic lion and snow leopard) in captive individuals (n = 8), of which 21 loci are being reported for the first time in the Bengal tiger, providing an additional choice for selection. The mean relatedness coefficient (R = -0.143) indicates that the selected tigers were unrelated. Thirty-four loci were polymorphic, with the number of alleles ranging from 2 to 7 per locus, and the remaining five loci were monomorphic. Based on the PIC values (> 0.500), and other characteristics, we suggest that 16 loci (3 to 7 alleles) be used for genetic and forensic study purposes. The probabilities of matching genotypes of unrelated individuals (3.692 × 10(-19)) and siblings (4.003 × 10(-6)) are within the values needed for undertaking studies in population genetics, relatedness, sociobiology and forensics.
    SpringerPlus 01/2014; 3(1):4. DOI:10.1186/2193-1801-3-4
  • Source
    • "Species identification was conducted using tiger-specific primers [19]. Based on expected heterozygosity, polymorphism and amplification success rates from previous studies in the laboratory, 14 microsatellite loci (FCA069, FCA090, FCA126, FCA230, FCA232, FCA279, FCA304, FCA441, FCA628, FCA672, [4], [18], [20], [21] MSHDZ170, MSFCA453, MSF115 and MSFCA506 [22] were used. PCR products were multiplexed and run with LIZ 500 size standard in an automated sequencer ABI3100XL (Applied Biosystems). "
    [Show abstract] [Hide abstract]
    ABSTRACT: Today, most wild tigers live in small, isolated Protected Areas within human dominated landscapes in the Indian subcontinent. Future survival of tigers depends on increasing local population size, as well as maintaining connectivity between populations. While significant conservation effort has been invested in increasing tiger population size, few initiatives have focused on landscape-level connectivity and on understanding the effect different landscape elements have on maintaining connectivity. We combined individual-based genetic and landscape ecology approaches to address this issue in six protected areas with varying tiger densities and separation in the Central Indian tiger landscape. We non-invasively sampled 55 tigers from different protected areas within this landscape. Maximum-likelihood and Bayesian genetic assignment tests indicate long-range tiger dispersal (on the order of 650 km) between protected areas. Further geo-spatial analyses revealed that tiger connectivity was affected by landscape elements such as human settlements, road density and host-population tiger density, but not by distance between populations. Our results elucidate the importance of landscape and habitat viability outside and between protected areas and provide a quantitative approach to test functionality of tiger corridors. We suggest future management strategies aim to minimize urban expansion between protected areas to maximize tiger connectivity. Achieving this goal in the context of ongoing urbanization and need to sustain current economic growth exerts enormous pressure on the remaining tiger habitats and emerges as a big challenge to conserve wild tigers in the Indian subcontinent.
    PLoS ONE 11/2013; 8(11):e77980. DOI:10.1371/journal.pone.0077980 · 3.23 Impact Factor
Show more