Abstract
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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Acknowledgements
This study was supported by the grant No. 90 preservation fund-2.1(8) from the COA and Department of Medical Research in NTUH, Taiwan, ROC.
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Supplementary Table 1
The primer sequences in this study. *The primer name is based upon the 3′ end of nucleotide site at cyt b gene sequence of Anderson et al. [24]. Supplementary material is available for this article at http://dx.doi.org/10.1007/s00414-XXX-XXX-X. (DOC 28 KB)
Supplementary Table 2
The primer pairs of nested PCR at cyt b gene in this study. a The fragment size is determined excluding the primer sequences for all PCR products and sequences upstream of the first codon on the cyt b gene. b The sample number is provided showing the number of samples successfully identified using one of the three strategies. Supplementary material is available for this article at http://dx.doi.org/10.1007/s00414-XXX-XXX-X. (DOC 38 KB)
Supplementary Table 3
The ranges of genetic distance illustrating interspecies and intraspecies variation. Supplementary material is available for this article at http://dx.doi.org/10.1007/s00414-XXX-XXX-X. (DOC 59 KB)
Supplementary Fig. 1
A neighbor-joining tree constructed from the sequence types of the three species of elephants using the 402 bp of the cyt b gene in this study. The bootstrap values at the nodes of the trees were obtained from 1,000 replicates and are shown as a percentage. Supplementary material is available for this article at http://dx.doi.org/10.1007/s00414-XXX-XXX-X. (DOC 118 KB)
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Lee, J.CI., Hsieh, HM., Huang, LH. et al. Ivory identification by DNA profiling of cytochrome b gene. Int J Legal Med 123, 117–121 (2009). https://doi.org/10.1007/s00414-008-0264-0
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DOI: https://doi.org/10.1007/s00414-008-0264-0