High-throughput collagen fingerprinting of intact microfaunal remains; a low-cost method for distinguishing between murine rodent bones.
High-throughput collagen fingerprinting of intact microfaunal remains; a low-cost method for distinguishing between murine rodent bones.
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DOI：
10.1002/rcm.7483
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发表时间：
2016-04-15
期刊：
影响因子：
--
通讯作者：
Chamberlain AT
中科院分区：
文献类型：
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作者：
Buckley M;Gu M;Shameer S;Patel S;Chamberlain AT
微型动物的骨骼遗骸可以作为当代生态系统的敏感指标，在这些生态系统中，它们被采样，并且经常在猫头鹰的颗粒中大量回收。这些遗骸的物种鉴定可以使用为特定骨骼元素建立的一系列形态学标准来获得，但通常依赖于颅骨特征。然而，在不同的环境和埋藏条件下，这可能会引起偏见。本研究的目的是开发一种高通量的方法，使用胶原蛋白指纹图谱客观地识别考古沉积物中的啮齿动物遗骸，最值得注意的是将大鼠从其他肌型啮齿动物中识别出来，作为通过存在与考古沉积物不同时的入侵类群来识别古动物群中干扰的一种手段。从完整的微型动物骨骼遗骸中提取胶原蛋白，以使骨骼形态完整（即，酸的浓度比以前使用的酸的浓度更低，时间更短）。然后将酸溶性胶原蛋白超滤至碳酸氢铵中，并用胰蛋白酶消化，然后在MALDI基质中稀释，并使用基质辅助激光解吸/电离飞行时间（MALDI-TOF）质谱仪采集肽质量指纹图谱。胶原蛋白指纹图谱在属水平上能区分鼠属、家鼠属、姬鼠属和田鼠属，在种水平上能区分鼠属和家鼠属。rattus和R. norvegicus可分离，A. flavicollis和A.西尔瓦提库斯不能。总共筛选了12，317个考古微脊椎动物样本的肌型特征，但没有发现侵入性大鼠（Rattus）或小鼠（Mus）。在当代鼠类区系中，没有发现收获鼠（Micromys），仅发现了24只野鼠（Apodemus）。因此，没有证据表明最近的生物扰动可以推断出这些考古沉积物的动物遗迹。更重要的是，这项工作提出了一种高通量筛选特定类群的方法，并且是胶原蛋白指纹图谱首次应用于考古标本的微型动物遗骸。版权所有2016作者.质谱快速通讯（Rapid Communications in Mass Spectrometry），John Wiley & Sons Ltd.
Microfaunal skeletal remains can be sensitive indicators of the contemporary ecosystem in which they are sampled and are often recovered in owl pellets in large numbers. Species identification of these remains can be obtained using a range of morphological criteria established for particular skeletal elements, but typically dominated by a reliance on cranial characters. However, this can induce biases under different environmental and taphonomic conditions. The aim of this research was to develop a high‐throughput method of objectively identifying rodent remains from archaeological deposits using collagen fingerprinting, most notably the identification of rats from other myomorph rodents as a means to identify disturbances in the archaeofauna through the presence of invasive taxa not contemporary with the archaeological deposits. Collagen was extracted from complete microfaunal skeletal remains in such a manner as to leave the bones morphologically intact (i.e., weaker concentration of acid than previously used over shorter length of time). Acid‐soluble collagen was then ultrafiltered into ammonium bicarbonate and digested with trypsin prior to dilution in the MALDI matrix and acquisition of peptide mass fingerprints using a matrix‐assisted laser desorption/ionisation time‐of‐flight (MALDI‐TOF) mass spectrometer. Collagen fingerprinting was able to distinguish between Rattus, Mus, Apodemus and Micromys at the genus level; at the species level, R. rattus and R. norvegicus could be separated whereas A. flavicollis and A. sylvaticus could not. A total of 12,317 archaeological microvertebrate samples were screened for myomorph signatures but none were found to be invasive rats (Rattus) or mice (Mus). Of the contemporary murine fauna, no harvest mice (Micromys) were identified and only 24 field mouse (Apodemus) discovered. As a result, no evidence of recent bioturbation could be inferred from the faunal remains of these archaeological deposits. More importantly this work presents a method for high‐throughput screening of specific taxa and is the first application of collagen fingerprinting to microfaunal remains of archaeological specimens. © 2016 The Authors. Rapid Communications in Mass Spectrometry Published by John Wiley & Sons Ltd.