Utility of DNA Barcoding for Identification of Bird-Strike Samples from India
Utility of DNA Barcoding for Identification of Bird-Strike Samples from India
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DNA 条形码在印度鸟击样品鉴定中的应用
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发表时间：
2016
期刊：
影响因子：
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通讯作者：
Y. Shouche
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作者：
Swapnil Gaikwad;H. Munot;Y. Shouche
鸟撞通常是由于飞机进入鸟层而发生的，鸟层是鸟类在天空中飞行的区域，在着陆或起飞时很常见。沿着经济损失，鸟撞还降低了机组人员的使命能力，损失飞行小时，对飞机造成永久性损害，重要的是，它总是与死亡率2-6的风险相关。减少鸟撞机会的策略包括改变生境、听觉和视觉的阻吓、鸟类雷达系统和改变飞机飞行时间和路线。为了实施这些策略，识别鸟撞中涉及的物种是重要的。但是，由于鸟撞的影响，鸟的身体被损坏得面目全非。通常，鸟撞击后的鸟的遗骸只包括血迹、羽毛或组织。基于羽毛形态学和角蛋白电泳的物种鉴定通常不能识别大多数报告的鸟撞案例2，7 -10。使用线粒体细胞色素c氧化酶I（COI）基因的DNA条形码已常规用于物种鉴定11。这种技术完全依赖于COI基因的核苷酸序列，因此可以从身体的任何部位进行物种鉴定，即使它已经损坏得无法识别。通常生成所述样品的COI基因序列并与COI基因序列数据库进行比较。全世界都在进行大规模的努力，利用DNA条形码12-15来鉴定鸟撞样本。据估计，印度航空业每年因鸟撞损失约300亿卢比。尽管它们对印度航空业产生了影响，但没有尝试使用DNA条形码来识别鸟撞样本。在本研究中，我们评估了DNA条形码的实用性，用于鸟撞事件中涉及的鸟类物种识别。从2011年到2014年，我们从印度海军收到了52个鸟撞样本。在大多数情况下，样本采集在Whatman FTA卡上（除非另有说明）。使用QIAamp ® DNA微型试剂盒（Qiagen，斯坦福大学，CA，USA），将一小片FTA卡用于DNA分离。第一次尝试未获得DNA的样品重复两次（表1）。COI基因的PCR扩增使用Dove等12中提到的引物进行。PCR产物经正向扩增后测序。
Bird strikes usually occur due to the entry of aircraft into the bird layer, i.e. the region up to which birds fly in the sky and are common during landing or takeoff 1. Along with the financial losses, bird strike also reduces the mission capability of the crew, loss of flying hours, permanent damages to the aircraft and importantly , it is always associated with the risk of mortality 2–6. The strategies to reduce the chances of bird strike involve habitat modifications, auditory and visual deterrents, avian radar system and changes in aircraft flight time and route. To implement these strategies, identification of the species involved in the bird strike is important. However, due to the impact of bird strike, the bodies of the birds are damaged beyond recognition. Usually the remains of the birds after a bird strike only include a smear of blood, feathers or tissues. Species identification based on feather morphology and keratin electrophoresis often fails to identify most of the reported bird-strike cases 2,7–10. DNA barcoding using mitochondrial Cytochrome c oxidase I (COI) gene has been routinely used for species identification 11. This technique depends entirely upon the nucleotide sequences of the COI gene and hence species identification is possible from any part of the body, even if it is damaged beyond recognition. Usually the COI gene sequence of the sample under question is generated and compared with the COI gene sequence database. All over the world large-scale efforts are being made to identify bird-strike samples using DNA barcoding 12–15. It is estimated that the aviation industry in India lose approximately Rs 30 billion/year due to bird strike 16. Despite their impact on the Indian aviation industry, no attempts are made to identify bird-strike samples using DNA barcoding. In the present study, we have evaluated the utility of DNA barcoding for species identification of birds involved in bird-strike incidences. We received 52 bird-strike samples from the Indian Navy from 2011 to 2014. In most of the cases, samples were collected on Whatman FTA cards (unless mentioned otherwise). A small piece of FTA card was utilized for DNA isolation using QIAamp ® DNA micro kit (Qiagen, Stanford, CA, USA). Samples for which DNA was not obtained in the first attempt were repeated twice (Table 1). PCR amplification of COI gene was done using primers mentioned by Dove et al. 12. The PCR products were sequenced using forward …