Recovering trace reptile DNA from the illegal wildlife trade

Nathan Deliveyne , Phillip Cassey , Adrian Linacre , Steven Delean , Jeremy J. Austin , Jennifer M. Young
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引用次数: 2

Abstract

Purpose

The Illegal Wildlife Trade (IWT), aided by improved global transport, and the expansion of the internet, has facilitated the international demand for exotic reptiles. The risks associated with trafficking of live reptiles requires robust forensic techniques for detecting housed or transported animals. Detection of species of high IWT demand can be challenging due to the illicit nature of the trade, particularly when a specimen is missing. The ability to detect trace DNA in empty holdings and transport containers can be pivotal as a source of evidence.

Methods

Vivaria, containing either a corn snake (Pantherophis guttatus) or boa (Boa constrictor), were set up and monitored for 24 h simulating reptile holdings. Once removed, Diamond Nucleic Acid Dye™ (DD) was sprayed on experimental glass and plastic tiles recovered from within the vivaria, and trace DNA was visualized using a Polilight. Trace DNA was amplified using a novel reptile target specific qPCR assay and sequenced to identify both species.

Results

Movement patterns and scale imprints associated with reptile contact were visible on experimental tiles after DD-staining. Successful qPCR amplification and subsequent bi-directional Sanger sequencing confirmed the presence of both the species in the respective vivaria. DNA recovered from glass tiles had significantly greater amplification success than plastic tiles.

Conclusions

DD revealed valuable information about the presence, and movement, of reptiles in the absence of a specimen. Successful amplification of trace reptile DNA demonstrated that this approach could offer an effective tool for biosecurity staff to rapidly identify live reptiles in the IWT.

从非法野生动物交易中提取爬行动物的DNA
目的非法野生动物贸易(IWT)在全球运输改善和互联网扩张的帮助下,促进了对外来爬行动物的国际需求。与贩运活爬行动物相关的风险需要强有力的法医技术来检测圈养或运输的动物。由于贸易的非法性质,特别是在标本丢失的情况下,检测内务运输需求高的物种可能具有挑战性。在空仓和运输容器中检测微量DNA的能力可能是关键的证据来源。方法设置含有玉米蛇(Pantherophis guttatus)或蟒蛇(boa constrictor)的vivaria,模拟爬行动物饲养,监测24 h。取出后,将钻石核酸染料™(DD)喷洒在从试管内回收的实验玻璃和塑料瓦上,并使用Polilight将痕量DNA可视化。使用一种新的爬行动物目标特异性qPCR扩增痕量DNA,并对其进行测序以鉴定这两个物种。结果经dd染色后,实验瓷砖上可见爬行动物接触的运动模式和鳞片印迹。成功的qPCR扩增和随后的双向Sanger测序证实了这两个物种在各自的种体内的存在。从玻璃瓦片中提取的DNA扩增成功率明显高于塑料瓦片。结论在标本缺失的情况下,ddd能提供有关爬行动物存在和运动的宝贵信息。痕量爬行动物DNA的成功扩增表明,该方法可以为生物安全人员提供快速识别内河野生动物活体的有效工具。
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来源期刊
Forensic science international. Animals and environments
Forensic science international. Animals and environments Pollution, Law, Forensic Medicine, Veterinary Science and Veterinary Medicine (General)
CiteScore
2.00
自引率
0.00%
发文量
0
审稿时长
142 days
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