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The Wartberg culture (WBC, 3500-2800 BCE) dates to the Late Neolithic period, a time of important demographic and cultural transformations in western Europe. We performed genome-wide analyses of 42 individuals who were interred in a WBC collective burial in Niedertiefenbach, Germany (3300-3200 cal. BCE). The results showed that the farming population of Niedertiefenbach carried a surprisingly large hunter-gatherer ancestry component (34–58%). This component was most likely introduced during the cultural transformation that led to the WBC. In addition, the Niedertiefenbach individuals exhibited a distinct human leukocyte antigen gene pool, possibly reflecting an immune response that was geared towards detecting viral infections., Alexander Immel et al. performed genome-wide analyses of 42 individuals from a collective burial in Niedertiefenbach, Germany from the Wartberg Culture. The authors find that this population had a large hunter-gatherer ancestry component and a distinct HLA pool, which indicates immune defenses against viral pathogens.

We studied HLA class I (HLA-A, -B) and class II (HLA-DRB1, -DQB1) alleles by PCR-SSP based typing in 1217 Mexicans from the Mexico City Metropolitan Area living in the northern (N = 751), southern (N = 52), eastern (N = 79), western (N = 33), and central (N = 152) Mexico City, and rural communities (N = 150), to obtain information regarding allelic and haplotypic frequencies. We found that the most frequent haplotypes include 11 Native American haplotypes. Admixture estimates revealed that the main genetic components are Native American (63.85 ± 1.55% by ML; 57.19% of Native American haplotypes) and European (28.53 ± 3.13% by ML; 28.40% of European haplotypes), and a less apparent African genetic component (7.61 ± 1.96% by ML; 7.17% of African haplotypes).

We combine the results of a radiocarbon (14C) dating program with archaeogenetic, osteological and sparse stratigraphic data, to construct a Bayesian chronological model for a multi-generational sequence situated entirely on a plateau in the 14C calibration curve. Calibrated dates of individual human bones from the Late Neolithic gallery grave at Niedertiefenbach, Hesse, Germany, span the entire calibration plateau in the late 4th millennium (ca. 3350–3100/3000 cal BC), but our model restricts the overall period of burial to 3–6 generations centered on the later 3200s, and provides narrower absolute date ranges for specific individuals and associated events. We confirm the accuracy and robustness of this model by sensitivity tests of each of its components. Beyond providing a more dynamic narrative for the formation of the heterogenous burial population at Niedertiefenbach, our results show that calibration plateaus are suitable periods for Bayesian chronological modeling of even relatively brief sequences, provided that all the information employed is correct. Prior information constraining both the order of events, and of potential date differences between them, is essential for the model to give accurate, unimodal estimates of the dates of these events.

The Cucuteni-Trypillia complex (CTC) flourished in eastern Europe for over two millennia (5100 – 2800 BCE) from the end of the Neolithic to the Early Bronze Age. Its vast distribution area encompassed modern-day eastern Romania, Moldova and western/central Ukraine. Due to a lack of existing burials throughout most of this time, only little is known about of the people associated with this complex and their genetic composition. Here, we present genome-wide data generated from the skeletal remains of four females that were excavated from two Late CTC sites in Moldova (3500 – 3100 BCE). All individuals carried a large Neolithic-derived ancestry component and were genetically more closely related to Linear Pottery than to Anatolian farmers. Three of the specimens also showed considerable amounts of steppe-related ancestry, suggesting influx into the CTC gene-pool from people affiliated with, for instance, the Ukraine Mesolithic. The latter scenario is supported by archaeological evidence. Taken together, our results confirm that the steppe component had arrived in eastern Europe farming communities maybe as early as 3500 BCE. In addition, they are in agreement with the hypothesis of ongoing contacts and gradual admixture between incoming steppe and local western populations.

Pathogens and associated outbreaks of infectious disease exert selective pressure on human populations, and any changes in allele frequencies that result may be especially evident for genes involved in immunity. In this regard, the 1346-1353 Yersinia pestis-caused Black Death pandemic, with continued plague outbreaks spanning several hundred years, is one of the most devastating recorded in human history. To investigate the potential impact of Y. pestis on human immunity genes we extracted DNA from 36 plague victims buried in a mass grave in Ellwangen, Germany in the 16th century. We targeted 488 immune-related genes, including HLA, using a novel in-solution hybridization capture approach. In comparison with 50 modern native inhabitants of Ellwangen, we find differences in allele frequencies for variants of the innate immunity proteins Ficolin-2 and NLRP14 at sites involved in determining specificity. We also observed that HLA-DRB1*13 is more than twice as frequent in the modern population, whereas HLA-B alleles encoding an isoleucine at position 80 (I-80+), HLA C*06:02 and HLA-DPB1 alleles encoding histidine at position 9 are half as frequent in the modern population. Simulations show that natural selection has likely driven these allele frequency changes. Thus, our data suggests that allele frequencies of HLA genes involved in innate and adaptive immunity responsible for extracellular and intracellular responses to pathogenic bacteria, such as Y. pestis, could have been affected by the historical epidemics that occurred in Europe. - Introduction - Results -- Archaeological and Anthropological Findings -- The 16th Century Ellwangen Plague Victims Display Genetic Similarity with Modern Inhabitants -- Two Immunity-Related Genes Harbor Strongly Differentiated single nucleotide polymorphisms -- No Evidence for Role of CCR5-D32 in Protection from Y. pestis Infection Discussion -- Natural Selection Has Increased HLA-DRB*13 and Reduced HLA-B*51 and -C*06 Frequencies in Modern Individuals -- Higher Incidence of KIR3DL1 Interaction with HLA-B in Plague Victims Than Modern Inhabitants of Ellwangen - Discussion - Materials and Methods

We studied HLA class I (HLA-A, -B) and class II (HLA-DRB1, -DQB1) alleles by PCR-SSP based typing in 81 Mexicans from the state of Campeche living in the city of Campeche (N = 34) and rural communities (N = 47), to obtain information regarding allelic and haplotypic frequencies. We find that the most frequent haplotypes in the state of Campeche include ten Native American, three European, one African and one Asian haplotype. Admixture estimates revealed that the main genetic components in the state of Campeche are Native American (65.56 ± 0.96% by ML; 51.24% of Native American haplotypes), European (34.44 ± 10.94% by ML; 30.25% of European haplotypes), and a virtually absent African genetic component (0.00 ± 10.31% by ML; 9.26% of African haplotypes).

It is a common assumption that – compared with the Mesolithic – the adoption of Neolithic lifeways was accompanied by a higher risk of infection and the development of epidemic diseases. Such a hypothesis seems plausible when considering singular archaeological parameters like increasing population density and palaeopathological indicators of poor health. However, evidence for the far-reaching consequences of epidemics has not yet been examined. Thus, the relevance of infectious diseases as triggers for transformation processes in the Neolithic remains to be identified. By reviewing specific archaeological, genetic and palaeopathological proxies gained from groups of individuals that inhabited the German loess zone from the early to the final Neolithic, we provide a diachronic view of the periods between 5500 and 2500 BCE with regard to postulated indicators of epidemic events. Our analyses of the archaeological proxies suggest major transformations in domestic strategies and mortuary practices, especially in the middle and late Neolithic. Interestingly, mass burials indicative of epidemic events are lacking. Ancient DNA results on pathogens confirm single infections throughout the Neolithic, but there is no clear evidence for diseases of epidemic proportions. The osteological records are not conclusive since the majority of osseous changes are unspecific with regard to the cause and course of infections. We conclude that currently neither biological nor archaeological proxies suggest substantial contributions of epidemics to Neolithic transformations in the German loess zone. This finding contrasts with the general assumption of a higher risk of infection and the development of epidemic diseases during the Neolithic.

In the Middle Ages, European settlements of all sizes were repeatedly affected by outbreaks of infectious diseases. Some of the outbreaks reached epidemic proportions, the second plague pandemic being the most notorious of all. A Late Medieval mass-burial site next to the Heiligen-Geist-Hospital (HGH) in Lubeck, a city of the Hanseatic League, contained the skeletal remains of more than 800 individuals assumed to have died of an infectious disease, most probably of the plague. However, Lubeck was ravaged by at least six pestilences in the 14 th century alone. We investigated ancient DNA extracts of 92 individuals from the HGH site to determine whether viral or bacterial pathogenic DNA was present that would help identify the cause of death. Metagenomic screening revealed evidence of an infection with Mi>Salmonella enterica subsp. enterica serovar Paratyphi C, suggesting an epidemic outbreak of enteric paratyphoid fever. A full reconstruction was possible for three S. Paratyphi C genomes that showed a close similarity to another strain from 1200 CE Norway. Based on radiocarbon dates, we determined the enteric paratyphoid fever outbreak in Lubeck to have occurred between 1360 and 1400 CE, historical records indicating the year 1367 CE as the most probable date. Our results also showed that the disease victims were people of northern and eastern European descent, confirming that Hanseatic Lubeck was an important trading centre for the Baltic region.

Summary A 5,000-year-old Yersinia pestis genome (RV 2039) is reconstructed from a hunter-fisher-gatherer (5300–5050 cal BP) buried at Riņņukalns, Latvia. RV 2039 is the first in a series of ancient strains that evolved shortly after the split of Y. pestis from its antecessor Y. pseudotuberculosis ∼7,000 years ago. The genomic and phylogenetic characteristics of RV 2039 are consistent with the hypothesis that this very early Y. pestis form was most likely less transmissible and maybe even less virulent than later strains. Our data do not support the scenario of a prehistoric pneumonic plague pandemic, as suggested previously for the Neolithic decline. The geographical and temporal distribution of the few prehistoric Y. pestis cases reported so far is more in agreement with single zoonotic events.

Ancient genomic studies have identified Yersinia pestis (Y. pestis) as the causative agent of the second plague pandemic (fourteenth–eighteenth century) that started with the Black Death (1,347–1,353). Most of the Y. pestis strains investigated from this pandemic have been isolated from western Europe, and not much is known about the diversity and microevolution of this bacterium in eastern European countries. In this study, we investigated human remains excavated from two cemeteries in Riga (Latvia). Historical evidence suggests that the burials were a consequence of plague outbreaks during the seventeenth century. DNA was extracted from teeth of 16 individuals and subjected to shotgun sequencing. Analysis of the metagenomic data revealed the presence of Y. pestis sequences in four remains, confirming that the buried individuals were victims of plague. In two samples, Y. pestis DNA coverage was sufficient for genome reconstruction. Subsequent phylogenetic analysis showed that the Riga strains fell within the diversity of the already known post-Black Death genomes. Interestingly, the two Latvian isolates did not cluster together. Moreover, we detected a drop in coverage of the pPCP1 plasmid region containing the pla gene. Further analysis indicated the presence of two pPCP1 plasmids, one with and one without the pla gene region, and only one bacterial chromosome, indicating that the same bacterium carried two distinct pPCP1 plasmids. In addition, we found the same pattern in the majority of previously published post-Black Death strains, but not in the Black Death strains. The pla gene is an important virulence factor for the infection of and transmission in humans. Thus, the spread of pla-depleted strains may, among other causes, have contributed to the disappearance of the second plague pandemic in eighteenth century Europe.

We studied HLA class I (HLA-A, -B) and class II (HLA-DRB1, -DQB1) alleles by PCR-SSP based typing in 112 Mexicans from the state of Morelos living in the city of Cuernavaca (N = 82) and rural communities (N = 30), to obtain information regarding allelic and haplotypic frequencies. The most frequent haplotypes in Morelos include seven Native American, one European, one African and one Asian haplotype. Admixture estimates revealed that the main genetic components in Morelos are Native American (60.43 ± 2.22% by ML; 53.57% of Native American haplotypes) and European (39.58 ± 3.70% by ML; 27.68% of European haplotypes), and a virtually absent African genetic component (0.00 ± 4.93% by ML; but 11.16% of African haplotypes).

We studied HLA class I (HLA-A, -B) and class II (HLA-DRB1, -DQB1) alleles by PCR-SSP based typing in 88 Mexicans from the state of Querétaro living in the city of Querétaro (N = 45) and rural communities (N = 43), to obtain information regarding allelic and haplotypic frequencies. We find that the most frequent haplotypes in the state of Querétaro include seven Native American, two European and one Asian haplotype. Admixture estimates revealed that the main genetic components in the state of Querétaro are Native American (51.82 ± 4.42% by ML; 42.61% of Native American haplotypes) and European (48.18 ± 3.55% by ML; 46.02% of European haplotypes), with a virtually absent African genetic component (0.00 ± 4.25% by ML; 4.55% of African haplotypes).

We studied HLA class I (HLA-A, -B) and class II (HLA-DRB1, -DQB1) alleles by PCR-SSP based typing in 173 Mexicans from the state of Chiapas living in the city of Tuxtla Gutiérrez (N = 52) and rural communities (N = 121), to obtain information regarding allelic and haplotypic frequencies. We found that the most frequent haplotypes in Chiapas include 12 Native American and one European haplotype. Admixture estimates revealed that the main genetic components in Chiapas are Native American (71.61 ± 0.58% by ML; 53.16% of Native American haplotypes) and European (26.39 ± 5.05% by ML; 25.86% of European haplotypes), and a less prominent African genetic component (2.00 ± 5.20% by ML; 9.77% of African haplotypes).

The Wartberg culture (WBC, 3,500-2,800 BCE) dates to the Late Neolithic period, a time of important demographic and cultural transformations in western Europe. We perform a genome-wide analysis of 42 individuals who were interred in a WBC collective burial in Niedertiefenbach, Germany (3,300-3,200 cal. BCE). Our results highlight that the Niedertiefenbach population indeed emerged at the beginning of the WBC. This farming community was genetically heterogeneous and carried a surprisingly large hunter-gatherer ancestry component (40%). We detect considerable differences in the human leukocyte antigen gene pool between contemporary Europeans and the Niedertiefenbach individuals whose immune response was primarily geared towards defending viral infections.

We studied HLA class I (HLA-A, -B) and class II (HLA-DRB1, -DQB1) alleles by PCR-SSP based typing in 122 Mexicans from the state of Hidalgo living in the city of Pachuca (N = 41) and rural communities (N = 81), to obtain information regarding allelic and haplotypic frequencies. We find that the most frequent haplotypes in Hidalgo include eight Native American and one European haplotypes. Admixture estimates revealed that the main genetic components in Hidalgo are Native American (58.93 ± 2.16% by ML; 54.51% of Native American haplotypes) and European (32.49 ± 2.88% by ML; 28.69% of European haplotypes), and a relatively high African genetic component (8.58 ± 0.93% by ML; 6.97% of African haplotypes).

We studied HLA class I (HLA-A, -B) and class II (HLA-DRB1, -DQB1) alleles by PCR-SSP based typing in 224 Mexicans from the state of Tabasco living in the city of Villahermosa (N = 82) and rural communities (N = 142), to obtain information regarding allelic and haplotypic frequencies. We found that the most frequent haplotypes in Tabasco include 13 Native American and two European haplotypes. Admixture estimates revealed that the main genetic components in Tabasco are Native American (67.79 ± 1.59% by ML; 56.25% of Native American haplotypes) and European (27.21 ± 3.97% by ML; 29.91% of European haplotypes), and a less prominent African genetic component (5.01 ± 4.42% by ML; 8.93% of African haplotypes).

The indigenous populations of inner Eurasia—a huge geographic region covering the central Eurasian steppe and the northern Eurasian taiga and tundra—harbour tremendous diversity in their genes, cultures and languages. In this study, we report novel genome-wide data for 763 individuals from Armenia, Georgia, Kazakhstan, Moldova, Mongolia, Russia, Tajikistan, Ukraine and Uzbekistan. We furthermore report additional damage-reduced genome-wide data of two previously published individuals from the Eneolithic Botai culture in Kazakhstan (~5,400 bp). We find that present-day inner Eurasian populations are structured into three distinct admixture clines stretching between various western and eastern Eurasian ancestries, mirroring geography. The Botai and more recent ancient genomes from Siberia show a decrease in contributions from so-called ‘ancient North Eurasian’ ancestry over time, which is detectable only in the northern-most ‘forest-tundra’ cline. The intermediate ‘steppe-forest’ cline descends from the Late Bronze Age steppe ancestries, while the ‘southern steppe’ cline further to the south shows a strong West/South Asian influence. Ancient genomes suggest a northward spread of the southern steppe cline in Central Asia during the first millennium bc. Finally, the genetic structure of Caucasus populations highlights a role of the Caucasus Mountains as a barrier to gene flow and suggests a post-Neolithic gene flow into North Caucasus populations from the steppe. Genome-wide data for 763 individuals from inner Eurasia reveal 3 admixture clines in present-day populations that mirror geography, illuminating the historic spread and mixture of peoples across the Eurasian steppe, taiga and tundra.

We studied HLA class I (HLA-A, -B) and class II (HLA-DRB1, -DQB1) alleles by PCR-SSP based typing in 144 Mexicans from the state of Guerrero to obtain information regarding allelic and haplotypic frequencies. We find that the ten most frequent haplotypes in the state of Guerrero include eight Native American and two European haplotypes. Admixture estimates revealed that the main genetic components in the state of Guerrero are Native American (61.36 ± 2.69% by ML; 54.17% of Native American haplotypes) and European (35.01 ± 4.59% by ML; 32.29% of European haplotypes), and a relatively low African genetic component (3.63 ± 2.38% by ML; 5.90% of African haplotypes).

The forced relocation of several thousand Africans during Mexico?s historic period has so far been documented mostly through archival sources, which provide only sparse detail on their origins and lived experience. Here, we employ a bioarchaeological approach to explore the life history of three 16th century Africans from a mass burial at the San José de los Naturales Royal Hospital in Mexico City. Our approach draws together ancient genomic data, osteological analysis, strontium isotope data from tooth enamel, δ13C and δ15N isotope data from dentine, and ethnohistorical information to reveal unprecedented detail on their origins and health. Analyses of skeletal features, radiogenic isotopes, and genetic data from uniparental, genome-wide, and human leukocyte antigen (HLA) markers are consistent with a Sub-Saharan African origin for all three individuals. Complete genomes of Treponema pallidum sub. pertenue (causative agent of yaws) and hepatitis B virus (HBV) recovered from these individuals provide insight into their health as related to infectious disease. Phylogenetic analysis of both pathogens reveals their close relationship to strains circulating in current West African populations, lending support to their origins in this region. The further relationship between the treponemal genome retrieved and a treponemal genome previously typed in an individual from Colonial Mexico highlights the role of the transatlantic slave trade in the introduction and dissemination of pathogens into the New World. Putting together all lines of evidence, we were able to create a biological portrait of three individuals whose life stories have long been silenced by disreputable historical events.

The indigenous populations of inner Eurasia, a huge geographic region covering the central Eurasian steppe and the northern Eurasian taiga and tundra, harbor tremendous diversity in their genes, cultures and languages. In this study, we report novel genome-wide data for 763 individuals from Armenia, Georgia, Kazakhstan, Moldova, Mongolia, Russia, Tajikistan, Ukraine, and Uzbekistan. We furthermore report genome-wide data of two Eneolithic individuals (∽5,400 years before present) associated with the Botai culture in northern Kazakhstan. We find that inner Eurasian populations are structured into three distinct admixture clines stretching between various western and eastern Eurasian ancestries. This genetic separation is well mirrored by geography. The ancient Botai genomes suggest yet another layer of admixture in inner Eurasia that involves Mesolithic hunter-gatherers in Europe, the Upper Paleolithic southern Siberians and East Asians. Admixture modeling of ancient and modern populations suggests an overwriting of this ancient structure in the Altai-Sayan region by migrations of western steppe herders, but partial retaining of this ancient North Eurasian-related cline further to the North. Finally, the genetic structure of Caucasus populations highlights a role of the Caucasus Mountains as a barrier to gene flow and suggests a post-Neolithic gene flow into North Caucasus populations from the steppe.

Riemerella anatipestifer is a Gram-negative bacterium belonging to the family Flavobacteriaceae . It is primarily associated with acute septicemia in younger birds. The R. anatipestifer isolate 17CS0503 described here was isolated from a Peking duck ( Anas platyrhynchos domesticus ) in Hannover, Germany, in 1999.

The hepatitis B virus (HBV) is one of the most widespread human pathogens known today, yet its origin and evolutionary history are still unclear and controversial. Here, we report the analysis of three ancient HBV genomes recovered from human skeletons found at three different archaeological sites in Germany. We reconstructed two Neolithic and one medieval HBV genomes byde novoassembly from shotgun DNA sequencing data. Additionally, we observed HBV-specific peptides using paleo-proteomics. Our results show that HBV circulates in the European population for at least 7000 years. The Neolithic HBV genomes show a high genomic similarity to each other. In a phylogenetic network, they do not group with any human-associated HBV genome and are most closely related to those infecting African non-human primates. These ancient virus forms appear to represent distinct lineages that have no close relatives today and went possibly extinct. Our results reveal the great potential of ancient DNA from human skeletons in order to study the long-time evolution of blood borne viruses.

Sub-fossilised remains may still contain highly degraded ancient DNA (aDNA) useful for palaeogenetic investigations. Whether X-ray computed [micro-] tomography ([μ]CT) imaging of these fossils may further damage aDNA remains debated. Although the effect of X-ray on DNA in living organisms is well documented, its impact on aDNA molecules is unexplored.Here we investigate the effects of synchrotron X-ray irradiation on aDNA from Pleistocene bones. A clear correlation appears between decreasing aDNA quantities and accumulating X-ray dose-levels above 2000 Gray (Gy). We further find that strong X-ray irradiation reduces the amount of nucleotide misincorporations at the aDNA molecule ends. No representative effect can be detected for doses below 200 Gy. Dosimetry shows that conventional μCT usually does not reach the risky dose level, while classical synchrotron imaging can degrade aDNA significantly. Optimised synchrotron protocols and simple rules introduced here are sufficient to ensure that fossils can be scanned without impairing future aDNA studies.

The giant deer Megaloceros giganteus is among the most fascinating Late Pleistocene Eurasian megafauna that became extinct at the end of the last ice age. Important questions persist regarding its phylogenetic relationship to contemporary taxa and the reasons for its extinction. We analyzed two large ancient cervid bone fragments recovered from cave sites in the Swabian Jura (Baden-Württemberg, Germany) dated to 12,000 years ago. Using hybridization capture in combination with next generation sequencing, we were able to reconstruct nearly complete mitochondrial genomes from both specimens. Both mtDNAs cluster phylogenetically with fallow deer and show high similarity to previously studied partial Megaloceros giganteus DNA from Kamyshlov in western Siberia and Killavullen in Ireland. The unexpected presence of Megaloceros giganteus in Southern Germany after the Ice Age suggests a later survival in Central Europe than previously proposed. The complete mtDNAs provide strong phylogenetic support for a Dama-Megaloceros clade. Furthermore, isotope analyses support an increasing competition between giant deer, red deer and reindeer after the Last Glacial Maximum, which might have contributed to the extinction of Megaloceros in Central Europe.