Development of PCR assay for differentiation of some important wild animal meat of Sri Lanka

A polymerase chain reaction (PCR) assay was developed to differentiate meat of Ceylon spotted deer (Axis axis ceylonensis), Ceylon hog deer (A. porcius oryzus), Ceylon sambhur (Cervus unicolor unicolor) and barking deer (Muntiacus muntijak malabaricus) from meat of cattle, goat, buffalo, pig, dog and sheep. A set of primers was designed according to the sequence of the mitochondrial cytochrome b gene of C. elaphus canadensis and by PCR amplification about 450 bp band was observed for all four animal species and these primers were not cross reacted with DNA of other animal species tested in the study under the tested reaction conditions. A band of 649 bp size was observed for all animal species when DNA was amplified with the universal primers and that indicated the presence of mitochondrial DNA in the samples. Further, the results indicated that this technique was sensitive enough to differentiate rotten meat, at least 5 days after the killing of an animal. Under these PCR conditions, the DNA of bacteria, which is involved in decomposition of meat, was not amplified with both universal and specific primers. However, the method was not sensitive enough in differentiating cooked meat of these species. Slaughtering of these four wild animal species is banned, but the animals are being killed illegally. Lack of meat identification methods has been identified as one of the major constraints to implement legal procedures and conserve biodiversity in the country.