Objective: To investigate the authenticity of infant Rhino horn sample from morphometric, microscopic, genetic and phylogenetic studies.
Methods:A suspected rhino horn sample (11 cm × 13 cm) was sent to the laboratory for identification. Relevant morphometric data of sample were taken. Similarly, established methods for microscopic examination of rhino horn were adopted. Using standard universal primer, a part of Cytochrome b (Cyt b) and 12S rRNA was amplified from sample, sequenced and aligned using BLAST of the GenBank (NCBI) and in-house reference database available at Wildlife Institute of India, Dehradun, India for decisive confirmation of species.Results: Morphometric analysis showed the sample was oval, dome shaped with a bony part which was supported by upper skin. Presence of filamentous units is the microscopic identifying feature of genuine rhino horn. However, microscopic examination showed very unclear filamentous units because of the immature nature of sample. Mitochondrial sequence data revealed the source of suspected sample was of a one-horn Rhinoceros (Rhinoceros unicornis). Similarly, NJ method for phylogenetic analysis for both Cytb and 12S rRNA identified sequence similarity with Rhinoceros unicornis.
Conclusion:Even though the morphometric and microscopic data do not reveal much about the infant rhino horn, mtDNA sequencing used in this pioneer study can be applied in the wildlife forensic for identification of such parts in future and data used as evidence against the poachers in the court of law.
As a part of wildlife crime investigation, two pieces of Asian elephant (Elephas maximus) ivory were collected by Uttarakhand forest department at different time interval to identify whether they belonged to same individual. The samples were analyzed using multilocus genotype based genetic analysis. Twelve fluorescent labeled microsatellite loci were used to analyze the nuclear DNA that were developed and validated for Asian elephants. Based on DNA typing, both the samples showed same alleles at every tested locus indicating that both the ivory pieces originated from single individual. The study highlights successful extraction and analysis of nuclear DNA employing microsatellite markers from ivory collected from Asian elephant.
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