DNA barcode based wildlife forensics for resolving the origin of claw samples using a novel primer cocktail

Abstract

Excessive wildlife hunting for commercial purposes can have negative impacts on biodiversity and may result in species extinction. To ensure compliance with legal statutes, forensic identification approaches relying on molecular markers may be used to identify the species of origin of animal material from hairs, claw, blood, bone, or meat. Using this approach, DNA sequences from the COI “barcoding” gene have been used to identify material from a number of domesticated animal species. However, many wild species of carnivores still present great challenges in generating COI barcodes using standard “universal” primer pairs. In the work presented here, the mitochondrial COI gene was successfully amplified using a novel primer cocktail, and the products were sequenced to determine the species of twenty one unknown samples of claw material collected as part of forensic wildlife case investigations. Sixteen of the unknown samples were recognized to have originated from either Panthera leo or P. pardus individuals. The remaining five samples could be identified only to the family level due to the absence of reference animal sequences. This is the first report on the use of COI sequences for the identification of P. pardus and P. leo from claw samples as part of forensic investigations in India. The study also highlights the need for adequate reference material to aid in the resolution of suspected cases of illegal wildlife harvesting.

Keywords:

• Claw
• cytochrome oxidase I gene
• DNA barcoding
• forensics
• India
• panthera
• primer cocktail

Acknowledgements

We are thankful to the Deputy Conservator of Forest, West Nasik Division for providing the samples for investigations. We acknowledge our lab colleagues for their support during laboratory work. The authors are thankful to Prof. David Haymer, Department of Cell and Molecular Biology, University of Hawaii for comments and suggestions on the draft manuscript.

Declaration of interest

All the authors declare that they have no competing interests. We are thankful to the Deputy Conservator of Forest, West Nasik Division for providing financial assistance to conduct the study