The rich fossil record of equids has made them a model for evolutionary processes. Here we present a 1.12-times coverage draft genome from a horse bone recovered from permafrost dated to approximately 560-780 thousand years before present (kyr BP). Our data represent the oldest full genome sequence determined so far by almost an order of magnitude. For comparison, we sequenced the genome of a Late Pleistocene horse (43 kyr BP), and modern genomes of five domestic horse breeds (Equus ferus caballus), a Przewalski's horse (E. f. przewalskii) and a donkey (E. asinus). Our analyses suggest that the Equus lineage giving rise to all contemporary horses, zebras and donkeys originated 4.0-4.5 million years before present (Myr BP), twice the conventionally accepted time to the most recent common ancestor of the genus Equus. We also find that horse population size fluctuated multiple times over the past 2 Myr, particularly during periods of severe climatic changes. We estimate that the Przewalski's and domestic horse populations diverged 38-72 kyr BP, and find no evidence of recent admixture between the domestic horse breeds and the Przewalski's horse investigated. This supports the contention that Przewalski's horses represent the last surviving wild horse population. We find similar levels of genetic variation among Przewalski's and domestic populations, indicating that the former are genetically viable and worthy of conservation efforts. We also find evidence for continuous selection on the immune system and olfaction throughout horse evolution. Finally, we identify 29 genomic regions among horse breeds that deviate from neutrality and show low levels of genetic variation compared to the Przewalski's horse. Such regions could correspond to loci selected early during domestication.

Calcified dental plaque (dental calculus) preserves for millennia and entraps biomolecules from all domains of life and viruses. We report the first high-resolution taxonomic and protein functional characterization of the ancient oral microbiome and demonstrate that the oral cavity has long served as a reservoir for bacteria implicated in both local and systemic disease. We characterize: (i) the ancient oral microbiome in a diseased state, (ii) 40 opportunistic pathogens, (iii) the first evidence of ancient human-associated putative antibiotic resistance genes, (iv) a genome reconstruction of the periodontal pathogen Tannerella forsythia, (v) 239 bacterial and 43 human proteins, allowing confirmation of a long-term association between host immune factors, “red-complex” pathogens, and periodontal disease, and (vi) DNA sequences matching dietary sources. Directly datable and nearly ubiquitous, dental calculus permits the simultaneous investigation of pathogen activity, host immunity, and diet, thereby extending the direct investigation of common diseases into the human evolutionary past.

One of the major difficulties in paleontology is the acquisition of fossil data from the 10% of Earth's terrestrial surface that is covered by thick glaciers and ice sheets. Here we reveal that DNA and amino acids from buried organisms can be recovered from the basal sections of deep ice cores and allow reconstructions of past flora and fauna. We show that high altitude southern Greenland, currently lying below more than two kilometers of ice, was once inhabited by a diverse array of conifer trees and insects that may date back more than 450 thousand years. The results provide the first direct evidence in support of a forested southern Greenland and suggest that many deep ice cores may contain genetic records of paleoenvironments in their basal sections.The environmental histories of high latitude regions such as Greenland and Antarctica are poorly understood because much of the fossil evidence is hidden below kilometer thick ice sheets (1-3). Here, we test the idea that the basal sections of deep ice cores can act as archives for ancient biomolecules and show that these molecules can be used to reconstruct significant parts of the past plant and animal life in currently ice covered areas.The samples studied come from the basal impurity rich (silty) ice sections of the 2km long Dye 3 core from south-central Greenland (4), the 3km long GRIP core from the summit of the UKPMC Funders Group Author Manuscript UKPMC Funders Group Author ManuscriptGreenland ice sheet (5), and the Late Holocene John Evans Glacier on Ellesmere Island, Nunavut, northern Canada (Fig. 1A,B). The latter sample was included as a control to test for potential exotic DNA because the glacier has recently overridden a land surface with a known vegetation cover (6). As an additional test for long-distance atmospheric dispersal of DNA, we included five control samples of debris-free Holocene and Pleistocene ice taken just above the basal silty samples from the Dye 3 and GRIP ice cores (Fig. 1B). Finally, our analyses included sediment samples from the Kap København Formation from the northernmost part of Greenland, dated to 2.4 million years before present (Ma BP) (1,2).The silty ice yielded only few pollen grains and no macrofossils (7). However, the Dye 3 and John Evans Glacier silty ice samples showed low levels of amino acid racemization (Fig. 1A, insert), indicating good organic matter preservation (8). Therefore, following previous success with permafrost and cave sediments (9-11), we attempted to amplify ancient DNA from the ice. This was done following strict criteria to secure authenticity (12-14), including covering the surface of the frozen cores with plasmid DNA to control for potential contamination that may have entered the interior of the samples through cracks or during the sampling procedure (7). PCR products of the plasmid DNA were obtained only from extracts of the outer ice scrapings but not from the interior, confirming that sample contamination had not penetrated the cores.We could reproducibly PCR amplify short ampli...