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      Barcoding the largest animals on Earth: ongoing challenges and molecular solutions in the taxonomic identification of ancient cetaceans

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          Abstract

          Over the last few centuries, many cetacean species have witnessed dramatic global declines due to industrial overharvesting and other anthropogenic influences, and thus are key targets for conservation. Whale bones recovered from archaeological and palaeontological contexts can provide essential baseline information on the past geographical distribution and abundance of species required for developing informed conservation policies. Here we review the challenges with identifying whale bones through traditional anatomical methods, as well as the opportunities provided by new molecular analyses. Through a case study focused on the North Sea, we demonstrate how the utility of this (pre)historic data is currently limited by a lack of accurate taxonomic information for the majority of ancient cetacean remains. We then discuss current opportunities presented by molecular identification methods such as DNA barcoding and collagen peptide mass fingerprinting (zooarchaeology by mass spectrometry), and highlight the importance of molecular identifications in assessing ancient species’ distributions through a case study focused on the Mediterranean. We conclude by considering high-throughput molecular approaches such as hybridization capture followed by next-generation sequencing as cost-effective approaches for enhancing the ecological informativeness of these ancient sample sets.

          This article is part of the themed issue ‘From DNA barcodes to biomes’.

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          Most cited references118

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          A draft sequence of the Neandertal genome.

          Neandertals, the closest evolutionary relatives of present-day humans, lived in large parts of Europe and western Asia before disappearing 30,000 years ago. We present a draft sequence of the Neandertal genome composed of more than 4 billion nucleotides from three individuals. Comparisons of the Neandertal genome to the genomes of five present-day humans from different parts of the world identify a number of genomic regions that may have been affected by positive selection in ancestral modern humans, including genes involved in metabolism and in cognitive and skeletal development. We show that Neandertals shared more genetic variants with present-day humans in Eurasia than with present-day humans in sub-Saharan Africa, suggesting that gene flow from Neandertals into the ancestors of non-Africans occurred before the divergence of Eurasian groups from each other.
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            Complete mitochondrial genome sequence of a Middle Pleistocene cave bear reconstructed from ultrashort DNA fragments.

            Although an inverse relationship is expected in ancient DNA samples between the number of surviving DNA fragments and their length, ancient DNA sequencing libraries are strikingly deficient in molecules shorter than 40 bp. We find that a loss of short molecules can occur during DNA extraction and present an improved silica-based extraction protocol that enables their efficient retrieval. In combination with single-stranded DNA library preparation, this method enabled us to reconstruct the mitochondrial genome sequence from a Middle Pleistocene cave bear (Ursus deningeri) bone excavated at Sima de los Huesos in the Sierra de Atapuerca, Spain. Phylogenetic reconstructions indicate that the U. deningeri sequence forms an early diverging sister lineage to all Western European Late Pleistocene cave bears. Our results prove that authentic ancient DNA can be preserved for hundreds of thousand years outside of permafrost. Moreover, the techniques presented enable the retrieval of phylogenetically informative sequences from samples in which virtually all DNA is diminished to fragments shorter than 50 bp.
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              The half-life of DNA in bone: measuring decay kinetics in 158 dated fossils.

              Claims of extreme survival of DNA have emphasized the need for reliable models of DNA degradation through time. By analysing mitochondrial DNA (mtDNA) from 158 radiocarbon-dated bones of the extinct New Zealand moa, we confirm empirically a long-hypothesized exponential decay relationship. The average DNA half-life within this geographically constrained fossil assemblage was estimated to be 521 years for a 242 bp mtDNA sequence, corresponding to a per nucleotide fragmentation rate (k) of 5.50 × 10(-6) per year. With an effective burial temperature of 13.1°C, the rate is almost 400 times slower than predicted from published kinetic data of in vitro DNA depurination at pH 5. Although best described by an exponential model (R(2) = 0.39), considerable sample-to-sample variance in DNA preservation could not be accounted for by geologic age. This variation likely derives from differences in taphonomy and bone diagenesis, which have confounded previous, less spatially constrained attempts to study DNA decay kinetics. Lastly, by calculating DNA fragmentation rates on Illumina HiSeq data, we show that nuclear DNA has degraded at least twice as fast as mtDNA. These results provide a baseline for predicting long-term DNA survival in bone.
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                Author and article information

                Journal
                Philos Trans R Soc Lond B Biol Sci
                Philos. Trans. R. Soc. Lond., B, Biol. Sci
                RSTB
                royptb
                Philosophical Transactions of the Royal Society B: Biological Sciences
                The Royal Society
                0962-8436
                1471-2970
                5 September 2016
                5 September 2016
                : 371
                : 1702 , Theme issue ‘From DNA barcodes to biomes’ compiled and edited by Paul D. N. Hebert, Mehrdad Hajibabaei and Peter M. Hollingsworth
                : 20150332
                Affiliations
                [1 ]BioArCh, Department of Archaeology, University of York , Environment Building, York, North Yorkshire YO10 5DD, UK
                [2 ]Institute of Archaeology, University College London , 31–34 Gordon Square, London WC1H 0PY, UK
                [3 ]CEFE UMR 5175, CNRS, Université de Montpellier, Université Paul-Valéry Montpellier, EPHE – CNRS , Montpellier Cedex 5, France
                [4 ]Archéologie des Sociétés Méditerranéennes, UMR 5140, CNRS, Labex Archimede IA-ANR-11-LABX-0032-01, Université Paul-Valéry Montpellier , 34970 Lattes, France
                [5 ]Dipartimento di Scienze della Natura e del Territorio, Università degli Studi , Sassari, Italy
                [6 ]Institute of Biochemistry and Biology, Faculty of Mathematics and Natural Sciences, University of Potsdam , 14476 Potsdam, Germany
                Author notes
                Author information
                http://orcid.org/0000-0003-0441-4705
                Article
                rstb20150332
                10.1098/rstb.2015.0332
                4971184
                27481784
                aa9efa05-6b5f-4297-a09c-36a8c834240c
                © 2016 The Authors.

                Published by the Royal Society under the terms of the Creative Commons Attribution License http://creativecommons.org/licenses/by/4.0/, which permits unrestricted use, provided the original author and source are credited.

                History
                : 28 April 2016
                Funding
                Funded by: French National Research Agency;
                Award ID: CEP&S 2011-ANR-279 11-CEPL-006
                Funded by: Labex Archimede programme;
                Award ID: IA- ANR-11-LABX-0032-01
                Funded by: European Research Council, http://dx.doi.org/10.13039/501100000781;
                Award ID: ORCA FP7-PEOPLE-2011-IIF 299075
                Categories
                1001
                70
                60
                197
                183
                144
                Articles
                Review Article
                Custom metadata
                September 5, 2016

                Philosophy of science
                ancient dna,archaeozoology,cetaceans,collagen peptide mass fingerprinting,species identification,zooarchaeology by mass spectrometry

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