Identification Of Smuggled Samples Of Genus Pygathrix

All materials which smuggled from Vietnam were captured by China customs, we must find out exact origins and species of these samples. The 28 dried samples belonged to genus Pygathrix, including hair, bones and muscle. This study based on mitochondrial control region HVâ… gene sequences, used sequence alignment with foreign gene bank of douc langur to identify the species and origins. Phylogenetic tree reconstructed by using neighbor-joining, minimum evolution, and maximum parsimony algorithm implemented in the computer software MEGA (molecular evolutionary genetic analysis) version 4.0. We used special primers did sex-typing. In order to study parentage of these 28 samples, we also used microsatellite DNA markers.In the 401bp sequences of control region, a total of 49 variable sites, 374 constant sites, 5 singleton sites, and 40 parsimony informative sites had been found. Among the sequences, there were 4 A-G and 6 T-C transitions sites on average, and more transitions than transversions. It bias against transitions, and has no gaps and insertions. The information of phylogenetic tree revealed the topology of phylogenetic tree constructed by the Neighbor-Joining method was accordant with that of the Maximum Parsimony tree and Minimum Evolution tree. NJ tree, ME tree and MP tree all primarily suggested the twenty-nine samples can divide into three clusters. According to the genetic pairwise distances results and compared with foreign gene bank information, we concluded 20 samples most likely originated from Khanh Hoa province in Vietnam. Another three came from Binh Phuoc province in Vietnam or Cambodian Mondulkiri province, whereas one sample came from Dak Lak or Lam Dong provinces, Vietnam. The origin of four further samples was not clearly traceable. Based on sex-typing PCRs, 17 samples are from females and 11 from males. This information indicated the 28 individuals maybe have some nature groups. We used 8 polymorphic microsatellite DNA primers to do paternity testing. The result showed there were several natural groups in Khanh Hoa samples. We concluded some samples from Khanh Hoa were poached by whole group; but other samples from different places were poached randomly.The smuggled samples were successfully identified by mtDNA sequences. This study provided a novel approach for the identification of douc langur samples in customs. Confirmed origins of smuggled samples were a strong support to protect the black-shanked Douc langur. Looking the map of Vietnam, we could easily find Khanh Hoa province facing to the South Sea of China, public transport was very convenience on the sea. Binh Phuoc province near by Cambodia closely, was one of the important main paths to Cambodia on the land. These advantageous geographic positions provided good conditions for the smuggler. After confirmed the origins, we could further observe illegal wildlife trade places and routes, provide trustworthy evidences to accurately beat smuggling activities of endanger Primates. This successful identification was due to international cooperation. The result of identification was important reference to international protection of douc langur. Furthermore, it was also significant for scientifically planning protection strategy of endanger Primates.