Unraveling the mystery of confiscated "jackal horns" in India using wildlife forensic tools.

Internationally, illegal wildlife trade involves highly prized and charismatic species and their derivatives. At the same time, common or less known species and their parts are also encountered but receive less attention than charismatic species. Given the increasing demand for wildlife products in many parts of the world, profit, and short supply, many fake articles derived from domestic or wild animals are frequently encountered in the wildlife trade. Jackal horn (locally known as "Siyar or Gidar singhi") is one such fake item widely used in sorcery and other occult practices available through offline and online trading platforms within India. We used a combination of morphological, microscopic hair, and molecular approaches (Cyt b and 16 s rRNA genes) to reveal the true identity of confiscated "jackal horns" (n = 342). Detailed morphological study of the jackal horns showed that it varied in size, shape, color of hair, attachment material, and filling material. The microscopic hair and molecular approaches revealed that all the items sold as jackal horns were fake and made up of protected wild species and domestic animals. Our results confirm the use of the biological samples from few wild species protected under the Wild Life (Protection) Act, 1972, of India. Therefore, the law enforcement agencies are cautioned to get forensic opinions while dealing with such counterfeit items.

Genetic portrait of North-West Indian population based on X chromosome Alu insertion markers.

In the present study, allele frequencies and forensic parameters of four ethnic groups (Brahmin, Khatri, Jat Sikh, and Scheduled Caste) of Punjab, India, at 10 Alu insertions of X chromosome were calculated. Six Alu markers were observed to be highly polymorphic with no significant deviations from Hardy-Weinberg equilibrium and no linkage disequilibrium present in any marker. Multidimensional plot showed higher genetic affinity of studied populations with Asian populations. Overall, the tested markers were reliable and were found suitable in human forensics and population genetic studies.

Genome Sequencing of a Gray Wolf from Peninsular India Provides New Insights into the Evolution and Hybridization of Gray Wolves.

The gray wolf (Canis lupus) is among the few large carnivores that survived the Late Pleistocene megafaunal extinctions. Thanks to their complex history of admixture and extensive geographic range, the number of gray wolf subspecies and their phylogenetic relationships remain poorly understood. Here, we perform whole-genome sequencing of a gray wolf collected from peninsular India that was phenotypically distinct from gray wolves outside India. Genomic analyses reveal that the Indian gray wolf is an evolutionarily distinct lineage that diverged from other extant gray wolf lineages ∼110 thousand years ago. Demographic analyses suggest that the Indian wolf population declined continuously decline since separating from other gray wolves and, today, has exceptionally low genetic diversity. We also find evidence for pervasive and mosaic gene flow between the Indian wolf and African canids including African wolf, Ethiopian wolf, and African wild dog despite their current geographical separation. Our results support the hypothesis that the Indian subcontinent was a Pleistocene refugium and center of diversification and further highlight the complex history of gene flow that characterized the evolution of gray wolves.

Morphometric variation in wolves and golden jackal in India (Mammalia, Carnivora).

Species of Canis (Carnivora, Canidae) have similar morphology and distinguishing sympatric species is challenging. We present data on morphometry of skull, body and hair of three wild Canis species that occur in India, which include two wolves (Indian wolf, Canislupuspallipes; and Himalayan wolf, Canishimalayensis) and the golden jackal (Canisaureus). A total of 20 cranial and six body measurements and microscopic characteristics of guard hair were analysed, using multivariate ordination to differentiate between species. Cranial measures of the Himalayan wolves were found to be the largest followed by Indian wolves and golden jackals. However, many measures overlapped amongst the three species. Two Principal Components each, for body measures and cranial measures, explained 86 and 91% of the variation in the data, respectively. These Components discriminated the two wolves from golden jackals, but could not distinguish between wolves. Hair medullary patterns were simple and wide type, whereas hair cuticular patterns showed crenate scale margins, near scale distance and irregular wavey scale patterns for all Canis taxa and were not useful to distinguish species. Data reported in this study further contribute to the existing global data on wild canids for a holistic understanding of the variation within the genus and show that distinguishing between all sympatric species from morphology alone may not be possible.

Identification of selected wild felids using hair morphology and forensically informative nucleotide sequencing (FINS): Wildlife forensics prospective.

We describe the identification of the leopard cat (Prionailurus bengalensis), jungle cat (Felis chaus) fishing cat (Prionailurus viverrinusis), wild cat (Felis silvestris) and caracal (Caracal caracal) using combined approach of morphological and molecular based analysis. Three mitochondrial genes [12S rRNA, 16S rRNA and cytochrome b (Cyt b)] and hair characteristics (cuticle and medulla patterns) were analysed and variation was observed among few selected wild felids. We did not find conclusive morphological difference among these examined felids. We observed maximum variation in Cyt b in comparison to12S rRNA and 16S rRNA genes. Therefore, despite poor hair morphological difference, forensically informative nucleotide sequencing (FINS) exhibited unambiguous variation among the examined felids. We suggest the use of FINS for differentiating biological samples of closely related wild felids to avoid any false identification of species in illicit trade. Furthermore, the data generated from the present work would help in strengthening the DNA database of Indian small cats.

Genetic variation and population structure of five ethnic groups from Punjab, North-West India: Analysis of MHC class I polymorphic Alu insertions (POALINs).

Genetic variation and differentiation of five ethnic groups from Punjab, North-West India was characterized by analyzing data on polymorphic Alu insertions (POALINs) within the class I genomic region of major histocompatibility complex (MHC), which is completely non-existent in Indian population. The haplotype frequency, distribution and heterozygosity among these groups and their potential implications in molecular anthropology and evolutionary studies were also determined. A total of 479 unrelated healthy individuals representing five different ethnic groups: Banias, Brahmins, Khatri, Jat Sikhs and Scheduled Castes were genotyped for five MHC Alu elements (AluHG, AluMICB, AluHJ, AluTF and AluHF) using polymerase chain reaction (PCR). All the loci were found to be polymorphic among the studied populations. No significant deviation from Hardy-Weinberg equilibrium was observed, except for the AluHJ locus in Brahmins. The POALINs varied in allele frequency between 0.0260 and 0.4427. The average heterozygosity among the studied groups ranged from 0.1937 in Banias to 0.2666 in Jat Sikhs. The genetic differentiation among the studied groups was observed to be of the order of 0.01302. Single POALIN haplotypes were found to be more frequent than multiple POALIN haplotypes. The results of inter-population differentiations, haplotype frequencies, genetic distances, multidimensional scaling, phylogenetic and structure analyses indicated close genetic relationships between the five ethnic groups of Punjab, North-West India. Analyses of polymorphic Alu loci of MHC genomic region may represent reliable information about the ancestry, demographic history and geographic origins of the various human populations, facilitating better understanding of the evolutionary, forensic and epidemiological prospective.