Ivory can be visually identified in its native form as coming from an elephant species; however, determining from which of the three extant elephant species a section of ivory originates is more problematic. We report on a method that will identify and distinguish the protected and endangered elephant species, Elephas maximus or Loxodonta sp. To identify the species of elephant from ivory products, we developed three groups of nested PCR amplifications within the cytochrome b gene that generate amplification products using highly degraded DNA isolated from confiscated ivory samples dating from 1995. DNA from a total of 382 out of 453 ivory samples were successfully isolated and amplified leading to species identification. All sequences were searched against GenBank and found to match with E. maximus and Loxodonta sp. with at least 99% similarity. The samples that were tested came from eight Asian elephants, 14 African forest elephants (Loxodonta cyclotis), and 360 African savannah elephants (Loxodonta africana). This study demonstrates a high success rate in species identification of ivory by a nested PCR approach within the cytochrome b gene which provides the necessary information for the protection of endangered species conservation.

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A DNA technique has been established for the identification to species level of tortoises. The test on the shell of the animal was used to identify samples from the species Kachuga tecta. A total of 100 tortoise shell specimens collected from the National Council of Agriculture (COA), Taiwan, were used in this study. Primer pairs were designed to amplify partial DNA fragments of cytochrome b within the mitochondrial genome. The DNA data showed that among the 100 samples, there were four distinct haplotype DNA sequences, within which there were a total of 90 variable sites. Between haplotypes I and II, there was only 1 nucleotide difference at position 228. Between haplotypes I and III, 65 nucleotide differences were observed; haplotypes I and IV, 62 nucleotide differences; and haplotypes III and IV, 56 nucleotide differences were observed. There were 66 and 63 nucleotide differences between haplotypes II and III and haplotypes II and IV respectively. All four haplotypes were compared with the DNA sequences held at the GenBank and EMBL databases. The most similar species were K. tecta (haplotype I and II), Morenia ocellata (haplotype III) and Geoclemys hamiltonii (haplotype IV), and their respective mtDNA similarities were 99.5%, 99.3%, 89.9% and 99.5%. However, as haplotype III was only 89.9% homologous with M. ocellata, it would seem that this haplotype shows only a limited relationship with a similar species registered currently in these databases. The method established by this study is an additional method for the identification of samples protected under Convention International Trade in Endangered Species (CITES) and will improve the work for the preservation of the endangered species.

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The population of the Tibetan Antelope (Pantholops hodgsonii) has recently declined dramatically due to the illegal trade in its wool. The animal lives at high altitude and is protected from the extremely cold climate due to a coat of very fine wool. These hairs are highly sought for weaving a shawl called shahtoosh. The large-scale poaching of the antelope has resulted in the species being placed on CITES Appendix I. We report on a method of DNA profiling on a confiscated shahtoosh using the cytochrome b (cyt b) gene to produce a test that will identify unambiguously the presence of P. hodgsonii. Two shahtoosh samples were provided by the Council of Agriculture, Taiwan, and ten shawl samples of sheep wool (Ovis aries), cashmere from the Kashmir goat (Capra hircus), and pashmina from the Himalayan goat (C. hircus) were collected from local stores for comparison. Two primer pairs were used to amplify a 271 bp fragment of cyt b gene which would distinguish these three species. The resulting amplification products were directly sequenced. When the DNA sequences were compared with the sequences registered in GenBank and EMBL databases, the sequences with the highest homology were the cyt b genes of P. hodgsonii, C. hircus, and O. aries. This study demonstrates that there is still sufficient DNA present in the finished wool of a shahtoosh shawl to allow DNA typing and the method established was highly plausible to identify the CITES protected species.