Aim: Chital and
sambar are the common wild small herbivores, which are vulnerable to poaching
for their meat. Many times poachers claim the wild meat to be that of goat or
sheep. Hence, authentic evidences are required to stop such wildlife crime. The
present investigation was carried out to study the species specific PCR-RFLP
patterns for meat identification of chital and sambar and then differentiation
from the meat of goat and sheep. Materials and Methods: Extracted DNA
from meat samples were subjected to PCR using the universal primers of 12S rRNA
gene. The PCR products were subjected to RFLP and sequencing.
The size of amplified PCR products was similar (440 bp) in each species and
sequence alignment showed more than 89 % similarities among these species.
However, phylogenetic analysis revealed that Chital and Sambar are in one
cluster while Goat and sheep are in other cluster. To differentiate between
species, restriction digestion of the PCR products was carried out to produce
characteristic PCR-RFLP patterns for each species. Restriction digestion with
RsaI and AluI enzymes produced distinct PCR-RFLP patterns that differentiated
the meat of wild species (chital-sambar) from that of domestic species
(goat-sheep). BsrI restriction digestion revealed unique PCR-RFLP pattern in
chital differentiating it from the meat of other three species. Restriction
digestion with DdeI enzyme led to the production of distinct PCR-RFLP patterns
for chital and sambar to identify their meat individually.
This study showed the effectiveness of 12S rRNA gene polymorphism in meat
identification. The data can be used as evidence against the poachers to convict
the wildlife crime in the court of law.
Keywords: 12S rRNA gene, meat
identification, restriction fragment length polymorphism (RFLP), wild small
herbivores, wildlife forensic