Discrimination of highly degraded, aged Asian and African elephant ivory using denaturing gradient gel electrophoresis (
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ORIGINAL ARTICLE
Discrimination of highly degraded, aged Asian and African elephant ivory using denaturing gradient gel electrophoresis (DGGE) Nitchakamon Suwanchatree 1 & Phuvadol Thanakiatkrai 1 & Adrian Linacre 2 & Thitika Kitpipit 1 Received: 8 May 2020 / Accepted: 27 August 2020 # Springer-Verlag GmbH Germany, part of Springer Nature 2020
Abstract Background Elephant populations have greatly reduced mainly due to illegal poaching for their ivory. The trade in elephant products is protected by national laws and CITES agreements to prevent them from further decline. For instance, in Thailand, it is illegal to trade ivory from African elephants; however, the law allows possession of ivory from Asian elephants if permission has been obtained from the authorities. As such, means of enforcement of legislation are needed to classify the legal status of seized ivory products. Many DNA-based techniques have been previously reported for this purpose, although all have a limit of detection not suitable for extremely degraded samples. Aim We report an assay based on nested PCR followed by DGGE to confirm the legal or illegal status of seized ivory samples where it is assumed that the DNA will be highly degraded. Method and results The assay was tested on aged ivory from which the assay was tested for reproducibility, specificity, and, importantly, sensitivity. Blind testing showed 100% identification accuracy. Correct assignment in all 304 samples tested was achieved including confirmation of the legal status of 227 highly degraded, aged ivories, thus underlining the high sensitivity of the assay. Conclusion and recommendation The research output will be beneficial to analyze ivory casework samples in wildlife forensic laboratories. Keywords DGGE . Ivory . Legal status identification
Introduction Elephants are large mammals of the Elephantidae family, of which there are three species currently recognized: the African bush elephant (Loxodonta africana), the African forest elephant (Loxodonta cyclotis), and the Asian elephant (Elephas maximus) [1–3]. All three elephant species have suffered significant population decline, with the main cause being deforestation which is their natural habitat; another reason is due to poaching and killing of elephants for their ivory or organs to be sold illegally [4–6]. Elephant species are now endangered species and are listed in Appendix I of the CITES Convention [7, 8], thus places a ban on
* Thitika Kitpipit [email protected]; [email protected] 1
Forensic Science Program, Department of Applied Science, Faculty of Science, Prince of Songkla University, 15 Karnjanavanich Road, Kho Hong, Hat Yai, Songkhla 90110, Thailand
2
College of Science and Engineering, Flinders University, Adelaide, SA 5042, Australia
international trade which strictly does not allow any import or export of any elephant products except for educational or research purpose. CITES restrictions do not affect the ownership and trade of ivory within a country. As an example, the law in Thailand allows possessio
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