For controlling illegal trade in wildlife parts and products in Southeast Asia, WII has initiated the work of establishing Wildlife Forensic Facility for proper implementation of Indian Wildlife (Protection) Act-1972. More than 500 wildlife offences have so far been referred by various agencies such as Forest, DRI, Courts, CBI, police and others. Most of the cases are of hair, wool, fur, skins, shawls, bones, claws, canine, antler, horn and others. In view of recent increase in number of meat related cases, we have established Wildlife Forensic DNA facility.

In the present talk, we discuss the initiatives undertaken for dealing meat related cases which can only be solved by DNA based techniques and characterization of four mongoose species based on hair characteristics. It has been globally accepted that there is a need for standardization of DNA extraction and PCR optimization protocols for the wildlife forensic materials, which are degraded and exposed to long-term environmental conditions. We tried three different methods for DNA extraction viz, phenol chloroform (PC), Chelex and commercial kits (Qaigen, Germany). The biological sample used for DNA extraction were hair, fresh tissue, blood, cooked meat, feathers, bear bile, musk pod & degraded meat. Successful DNA extraction from blood, meat and hair samples were in order of Qaigen>Chelex>PC whereas in case of hair samples, PC as well as Chelex were satisfactorily good.

During PCR amplification (n=184), 53% of DNA from these samples did not amplify. Of the remaining amplified samples (n=97), 47% resulted in good amplification (++), 35% gave faint product (+), and 8% very faint (-). The PCR amplification success rate with conserved regions of vertebrate mitochondrial cytochrome b gene (359 bp) using DNA extracted from sambar, chital and mongoose by various methods was in the order of Qaigen>PC>Chelex.

We used six restriction enzymes viz,.Hae III, Alu I,Rsa I,Msp I,EcoR I and Hind III and the success rate of restriction digestion in relation to the above mentioned extracted methods was in the order of Qaigen (90%) > Chelex (75%) > PC (55%). Data on Sambar (Cervus unicolor), Chital (Axis axis) and mongoose indicates that it is possible to differentiate these species using the restriction enzymes Hae III, Msp I and Rsa I. Another DNA technique RAPD (Random amplified Polymorphic DNA) was tried for 17 samples and success in RAPD from samples extracted by various DNA methods was in the order of Qaigen (75%) > PC(25%) > Chelex(0%).