Abstract
Wolf (Canis lupus) is a species included in appendices of CITES and is often encountered in cases of alleged poaching and trafficking of their products. When such crimes are suspected, those involved may attempt to evade legal action by claiming that the animals involved are domestic dogs (C. l. familiaris). To respond effectively to such claims, law enforcement agencies require reliable and robust methods to distinguish wolves from dogs. Reported molecular genetic methods are either unreliable (mitogenome sequence based), or operationally cumbersome and require much DNA (un-multiplexed microsatellites), or financially expensive (genome wide SNP genotyping). We report on the validation of a panel of 12 ancestral informative single nucleotide polymorphism (SNP) markers for discriminating wolves from dogs. A SNaPshot multiplex genotyping system was developed for the panel, and 97 Mongolian wolves (C. l. chanco) and 108 domestic dogs were used for validation. Results showed this panel had high genotyping success (0.991), reproducibility (1.00) and origin assignment accuracy (0.97 ± 0.05 for dogs and 1.00 ± 0.03 for wolves). Species-specificity testing suggested strong tolerance to DNA contamination across species, except for Canidae. The minimum DNA required for reliable genotyping was 6.25 pg/μl. The method and established gene frequency database are available to support identification of wolves and dogs by law enforcement agencies.
Highlights
Wolf (Canis lupus) is a species included in appendices of CITES and is often encountered in cases of alleged poaching and trafficking of their products
In attempts to avoid penalties, traders often claimed that the bones and canines were those of large dogs and were sold as counterfeits of wolf body parts
Identification based on control region (CR) sequence may not be reliable unless additional nuclear markers are combined[32,33] especially when the reference sample size is not sufficient to represent total genetic v ariation[34]
Summary
Wolf (Canis lupus) is a species included in appendices of CITES and is often encountered in cases of alleged poaching and trafficking of their products When such crimes are suspected, those involved may attempt to evade legal action by claiming that the animals involved are domestic dogs In attempts to avoid penalties, traders often claimed that the bones and canines were those of large dogs and were sold as counterfeits of wolf body parts Prosecution of these cases is dependent on definitive subspecies identification. Familiaris[19], but their gene pools exhibit great homogeneity and contain detectable historical and recent genetic admixtures[20,21,22,23,24] For this reason, coding regions of the mitochondrial genome have either no resolution between wolf and dog (12S and 16S ribosomal RNA genes)[25,26], or detectable but poor resolution (Cyt b and COI genes)[27]. The results have become a rich and convenient resource for forensic identification of wolf and dog
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