First direct dating of the Late Neanderthal remains from Subalyuk Cave in Northern Hungary.

The Subalyuk hominin remains were uncovered in 1932 in a cave of the same name in the Bükk Mountains, near the village of Cserépfalu in Borsod-Abaúj-Zemplén County, Northern Hungary. The remains represent two individuals, an adult and a young child who have been described in a few publications since their discovery, providing substantial anthropological data and general assessments of their Neanderthal affiliation. They were associated with Late Mousterian industry. Thus, the Bükk Mountains gain importance in the discussion concerning the contribution of East Central European sites to the debate on the peopling history of Europe during the Late Middle to Early Upper Palaeolithic transition. In this paper, we summarize the archaeological and chronological context of the two individuals, and publish the first direct dating results that place them among the Last Neanderthals of Central Europe.

The paleopathology and paleoepidemiology of Upper paleolithic tuberculosis: Review of evidence and hypotheses.

Molecular phylogeny work has shown that tuberculosis is ancient human-adapted infection predating the Neolithic period. They also show that the Upper Paleolithic is a key period of emergence of the MTB complex strains, contemporary with the exit of modern man from Africa. Despite the richness of Upper Paleolithic sites in Eurasia and the relative abundance of human remains, the only proven case of Paleolithic tuberculosis has been described so far date from the Azilian, a culture of the European Final Paleolithic, which is more recent than the ancient Neolithic sites of the Near East, area that currently hold the record for the oldest paleopathological evidence of tuberculosis. The purpose of this review is to present evidence for the existence of tuberculosis in the Paleolithic and to list hypotheses explaining the weak demonstrative contribution of paleopathology for pre-Neolithic periods.

Oldest evidence of tuberculosis in the Mediterranean islands: From the mainland to Cyprus.

Recent studies combining macroscopical observations and microCT analysis strongly suggested the diagnosis of tuberculosis for a child from the site of Khirokitia (Cyprus, 7th - early 6th millennium cal. BC), whose age at death is between 5 and 7 years. Many single primary burials were discovered at the site where the dead (MNI = 243) are buried in the same way, whatever their age. Nevertheless, the burial of this child presents a unique feature on the site (a male Ovis trophy marking the limit of the burial pit), probably indicating specific attention for this young deceased. This case is the oldest known in the Mediterranean islands and presents a particular interest from a paleoepidemiological point of view. Indeed, considering, on the one hand, the settlement pattern of the island of Cyprus by migrants from the Near East, and on the other hand, the presence of human tuberculosis in the Near East as early as about 10,500 years BP, it is very likely that the prehistoric migrants brought the disease from mainland to Cyprus.

Re-examination of the Subalyuk Neanderthal remains uncovers signs of probable TB infection (Subalyuk Cave, Hungary).

In 1932, skeletal remains of two Neanderthal individuals, a young adult female and a 3-4-year-old child, were discovered in Subalyuk Cave in Northern Hungary [1,2]. Results of the anthropological examination were published some years after this important discovery. Methodological progress encouraged re-examination of the material during the last few years. Radiocarbon dating revealed a chronological age of 39,732-39,076 cal. BP for the adult female and 36,117-35,387 cal. BP for the child [3]. Morphological paleopathological studies of these Neanderthal remains uncovered distinct evidence of skeletal infections. Alterations of the adult individual's sacrum suggest probable early-stage sacroiliitis, while several vertebral bodies indicate superficial osseous remodelling of infectious origin. Traces of pathological lesions were observed on the endocranial surface of the child's skull, reflecting a reaction of meningeal tissues, a consequence of a probable TB-related meningeal infectious process. Results of recent paleomicrobiological examinations - lipid biomarker and aDNA studies - support the morphological diagnosis of probable TB infections [4].

Upper Paleolithic tuberculosis: A probable case illustrated by paleoimaging methods (Azilian site of Les Iboussières, France).

The aim of this paper is to present the results of µCT-scan and 3D imaging analyses of two skeletal lesions observed on human remains of one of the last European hunter-gatherers from the late Paleolithic (Azilian period): a sacroiliac osteoarthritis and a femoral lesion suggestive of a soft tissue abscess imprint. These two skeletal elements (fused left sacrum and coxal bone, and right femur) displayed osteometric criteria indicating that they belonged to the same individual. These two associated lesions are consistent with a low-grade osteoarticular infection, and suggest a diagnosis of pelvic tuberculosis with a cold abscess of the thigh. If molecular confirmation is obtained, this case would be the first evidence of tuberculosis among Upper Palaeolithic populations.

Inner ear modifications in Dar-es-Soltane II H5 (Morocco): A case of labyrinthitis ossificans.

OBJECTIVE: This paper presents the inner ear modifications in Dar-es-Soltane II H5, an Aterian fossil possibly dated to 100 ka. MATERIAL: The remains consist of a large portion of the cranium including the face, the left frontal and temporal bones, part of the left parietal bone and greater wing of the sphenoid. METHODS: The bony labyrinth anatomy was investigated on existing micro-CT data acquired by the MPI-EVA. RESULTS: The observation of micro-CT sections revealed a partial filling of the semi-circular canals that raises question about its origin. A careful examination of the micro-CT sections shows that the elements present in the semicircular canals were denser than the sediments observed in other regions and cavities of the temporal bone. CONCLUSIONS: The current evidence suggests a pathological origin of this condition with partial ossification of the membranous labyrinth. The differential diagnosis indicates a case of labyrinthitis ossificans in its early stages. SIGNIFICANCE: This pathological condition can be responsible for permanent hearing loss and is associated with dizziness and vertigo. Along with the Singa skull, Dar-es-Soltane II H5 represents one of the oldest known cases of labyrinthitis ossificans. LIMITATIONS: The early stage of disease and the absence of the right temporal bone limit conclusions about the degree of disability of the individual and their dependence on the rest of the group. SUGGESTIONS FOR FURTHER RESEARCH: To carry out a paleopathological study of all the fossils from Dar-es-Soltane II.

Can the palaeoepidemiology of rickets during the industrialisation period in France be studied through bioarchaeological grey literature and French medico-historical literature of the 18th-early 20th centuries? Preliminary examination of a complex topic.

OBJECTIVE: This study explores whether data relating to rickets from the French medico-historical literature (FMHL) and bioarchaeological grey literature are useful in evaluating its epidemiology during the industrialisation of France. Unlike other European countries such as England, industrialisation in France was a slow and continuous process with two phases: the first in 1830-1870 and the second in 1870-1914. MATERIALS AND METHODS: A bibliographical analysis of 2800 FMHL sources from the 18th to the early 20th centuries and 50 archaeological excavation reports from the last 21 years was undertaken. RESULTS: The FMHL data is very heterogeneous and predominantly dates to the second phase of industrialisation. The bioarchaeological data is very incomplete and predominantly relates to the period before industrialisation. At the same time, knowledge improvement and institutional changes to protect children could explain more systematic registration of cases of rickets. CONCLUSIONS: No solid conclusions can be made regarding the prevalence of rickets at present, however these data hold great potential. SIGNIFICANCE: In comparison to England, no systematic investigation of rickets prevalence during the period of industrialisation in France has been undertaken to date. LIMITATIONS: The lack of archaeological excavations from this period and the limited paleopathological analysis of the sites excavated have contributed to our current lack of understanding regarding the impact of industrialization on the prevalence of rickets on the French population. SUGGESTIONS FOR FURTHER WORK: The FMHL data needs to be homogenized and osteoarchaeological collections need to be restudied with a common protocol focusing on signs of vitamin D deficiency.

Was the rise of TB contemporaneous with the industrial revolution? Epidemiological evolution of TB in France (17th-20th centuries) inferred from osteoarchaeological and historical archives.

OBJECTIVE: The development of tuberculosis is classically associated with the rise of the Industrial Revolution. Our objective is to test this epidemiological hypothesis for populations in France between the 18th and 20th centuries using osteoarchaeological and historical sources. MATERIALS: Osteoarchaeological sources include two skeletal collections from plague epidemics in Provence (1590 and 1722) representing a total of 349 individuals and medical archives dating 1750-1930 from Paris, Lyon, Marseille, Montpellier. METHODS: Paleoepidemiological analysis (crude prevalence rate) of the archeological data, and epidemiological analysis of historical data included the proportional mortality rate of tuberculosis (PMR-TB) and the mortality rate by tuberculosis (MR-TB). RESULTS: Mean prevalence of TB before the 19th century was approximately 30 %, according to osteoarchaeological data. Historical sources showed that pulmonary TB was responsible for 33 % of total deaths in the city of Marseille during the second half of the 18th century and represented about 20 % of deaths in the four French cities during the 19th century. The mortality rate was 6.5 per thousand inhabitants in the 18th century (Marseille) and stable at 4-5 ‰ during the 19th century before, during, and after the Industrial Revolution period, with the exception of years 1870-1871 (French-Prussian war), when it increased. CONCLUSIONS: Population increase contemporary to the industrialization process did not increase the mortality rate by tuberculosis in France. SIGNIFICANCE: The epidemiological assertion that tuberculosis increased with the Industrial Revolution in cities must be reevaluated. In France, and perhaps in other cities, it was an endemic disease at least a century before, and associated with a higher mortality rate. LIMITATIONS: Reliability and biases inherent to archaeological and historical data prior and during the Industrial Revolution must be taken into account. SUGGESTIONS FOR FURTHER RESEARCH: Extension of research to all the French territories (rural and urban areas) is advised.

Are the identified collections of immature skeletons dating from the Industrial Revolution good references for paleoauxological studies? Cases studies from England & France.

OBJECTIVE: Skeletal collections of immature individuals identified by age and sex serve as reference material for studying development in past populations. Several of these collections were established during the Industrial Revolution (IR), a period known for its difficult living conditions in industrial cities. We question if these collections represent useful comparisons from which to explore the natural history of human growth. MATERIALS: Immature individuals from two skeletal collections contemporaneous to the IR period were studied: 71 children from the Spitalfields (UK) and 108 from the Strasbourg (F) collections. Among them we selected mandibles of individuals aged from 0 to 30 months, representing 32 and 52 individuals, respectively. METHODS: We scored the dental development of first and second left deciduous molars according to (Moorrees et al., 1963) stages, from X-rays (Spitalfields) or CT-scans (Strasbourg) data and compared it with the modern reference pattern from the Lewis Growth Records by covariance analysis (ANCOVA). RESULTS: Statistical differences exist in the dental development timeline between the 3 samples. This mainly concerns a delay in the root formation in IR samples that related to post-natal living conditions. CONCLUSIONS: The delay in dental development timeline suggests that growth processes were impacted during IR in England and France, probably due to stressful living conditions. SIGNIFICANCE: keletal collections dating from the IR period in Europe might be not the most appropriate referencesl for studying the natural history of human growth. LIMITATIONS: This study focus on dental development only. SUGGESTIONS FOR FURTHER RESEARCH: Exploring the skeletal growth pattern in other skeletal collections, pre or post-dating the IR, is advised.

A unique case of skeletal dysplasia in an adult male in Late Iron Age Switzerland.

OBJECTIVE: We report a probable case of multiple skeletal dysplasia observed in a Late Iron Age young adult male. MATERIALS: The individual studied belongs to a Late Iron Age necropolis from Switzerland. The skeletal elements are well preserved METHODS: Macroscopic and radiographic assessment. RESULTS: The individual shows evidence of both craniofacial and mandibular deformation. Developmental defects are also visible with effects on the general shape and articular surfaces of both humeri, as well as the left femur and tibia. CONCLUSION: We propose that the lesions observed are manifestations of skeletal dysplasia, such as pseudo-achondroplasia or multiple epiphyseal dysplasia. SIGNIFICANCE: This is the first recorded case of multiple skeletal dysplasia in an Iron Age necropolis in Switzerland, questioning the integration of physically compromised individuals in La Tène society. LIMITATIONS: Examination of other skeletal dysplasias from archaeological contexts provides support for this diagnosis. SUGGESTIONS FOR FUTURE RESEARCH: The integration of individuals with disabilities in La Tène societies is still poorly understood and further research is needed to better characterize these communities.

Five millennia of Bartonella quintana bacteraemia.

During the two World Wars, Bartonella quintana was responsible for trench fever and is now recognised as an agent of re-emerging infection. Many reports have indicated widespread B. quintana exposure since the 1990s. In order to evaluate its prevalence in ancient populations, we used real-time PCR to detect B. quintana DNA in 400 teeth collected from 145 individuals dating from the 1st to 19th centuries in nine archaeological sites, with the presence of negative controls. Fisher's exact test was used to compare the prevalence of B. quintana in civil and military populations. B. quintana DNA was confirmed in a total of 28/145 (19.3%) individuals, comprising 78 citizens and 67 soldiers, 20.1% and 17.9% of which were positive for B. quintana bacteraemia, respectively. This study analysed previous studies on these ancient samples and showed that the presence of B. quintana infection followed the course of time in human history; a total of 14/15 sites from five European countries had a positive prevalence. The positive rate in soldiers was higher than those of civilians, with 20% and 18.8%, respectively, in the 18th and 19th centuries, but the difference in frequency was not significant. These results confirmed the role of dental pulp in diagnosing B. quintana bacteraemia in ancient populations and showed the incidence of B. quintana in both civilians and soldiers.

Trabecular bone microarchitecture analysis, a way for an early detection of genetic dwarfism? Case study of a dwarf mother's offspring.

A 66 year-old woman with a disproportionate dwarfism and who bore seven children was discovered at the Middenbeemster archaeological site (The Netherlands). Three are perinates and show no macroscopic or radiological evidence for a FGFR3 mutation causing hypo-or achondroplasia. This mutation induces dysfunction of the growth cartilage, leading to abnormalities in the development of trabecular bone. Because the mutation is autosomal dominant, these perinates have a 50% risk of having been affected. This study determines whether trabecular bone microarchitecture (TBMA) analysis is useful for detecting genetic dwarfism. Proximal metaphyses of humeri were µCT-scanned with a resolution of 7-12 µm. Three volumes of interest were segmented from each bone with TIVMI© software. The TBMA was quantified in BoneJ© using six parameters on which a multivariate analysis was then performed. Two of the Middenbeemster perinates show a quantitatively different TBMA organization. These results and the family's medical history suggest a diagnosis of genetic dwarfism for this two perinates. This study provides evidence to support the efficacy of µCT for diagnosing early-stage bone disease.

17th Century Variola Virus Reveals the Recent History of Smallpox.

Smallpox holds a unique position in the history of medicine. It was the first disease for which a vaccine was developed and remains the only human disease eradicated by vaccination. Although there have been claims of smallpox in Egypt, India, and China dating back millennia [1-4], the timescale of emergence of the causative agent, variola virus (VARV), and how it evolved in the context of increasingly widespread immunization, have proven controversial [4-9]. In particular, some molecular-clock-based studies have suggested that key events in VARV evolution only occurred during the last two centuries [4-6] and hence in apparent conflict with anecdotal historical reports, although it is difficult to distinguish smallpox from other pustular rashes by description alone. To address these issues, we captured, sequenced, and reconstructed a draft genome of an ancient strain of VARV, sampled from a Lithuanian child mummy dating between 1643 and 1665 and close to the time of several documented European epidemics [1, 2, 10]. When compared to vaccinia virus, this archival strain contained the same pattern of gene degradation as 20th century VARVs, indicating that such loss of gene function had occurred before ca. 1650. Strikingly, the mummy sequence fell basal to all currently sequenced strains of VARV on phylogenetic trees. Molecular-clock analyses revealed a strong clock-like structure and that the timescale of smallpox evolution is more recent than often supposed, with the diversification of major viral lineages only occurring within the 18th and 19th centuries, concomitant with the development of modern vaccination.

Paleopathology of Human Infections: Old Bones, Antique Books, Ancient and Modern Molecules.

Paleopathology studies the traces of disease on human and animal remains from ancient times. Infectious diseases have been, for over a century, one of its main fields of interest. The applications of paleogenetics methods to microbial aDNA, that started in the 90s combined to the recent development of new sequencing techniques allowing 'paleogenomics' approaches, have completely renewed the issue of the infections in the past. These advances open up new challenges in the understanding of the evolution of human-pathogen relationships, integrated in "One Health" concept.In this perspective, an integrative multidisciplinary approach combining data from ancient texts and old bones to those of old molecules is of great interest for reconstructing the past of human infections. Despite some too optimistic prediction of their eradication in the late 20th century, some of these ancient human diseases, such as plague, leprosy or tuberculosis, are still present and continue their evolution at the beginning of this 21rst century. Better know the past to predict a part of the future of human diseases remains, more than ever, the motto of the paleopathological science.

Eighteenth century Yersinia pestis genomes reveal the long-term persistence of an historical plague focus.

The 14th-18th century pandemic of Yersinia pestis caused devastating disease outbreaks in Europe for almost 400 years. The reasons for plague's persistence and abrupt disappearance in Europe are poorly understood, but could have been due to either the presence of now-extinct plague foci in Europe itself, or successive disease introductions from other locations. Here we present five Y. pestis genomes from one of the last European outbreaks of plague, from 1722 in Marseille, France. The lineage identified has not been found in any extant Y. pestis foci sampled to date, and has its ancestry in strains obtained from victims of the 14th century Black Death. These data suggest the existence of a previously uncharacterized historical plague focus that persisted for at least three centuries. We propose that this disease source may have been responsible for the many resurgences of plague in Europe following the Black Death.

Tuberculosis in Late Neolithic-Early Copper Age human skeletal remains from Hungary.

Alsónyék-Bátaszék in Southern Hungary is one of the largest late Neolithic settlements and cemeteries excavated in Central Europe. In total, 2359 burials from the Late Neolithic - Early Copper Age Lengyel culture were found between 2006 and 2009 [1]. Anthropological investigations previously carried out on individuals from this site revealed an interesting paleopathological case of tuberculosis in the form of Pott's disease dated to the early 5(th) millennium BC. In this study, selected specimens from this osteoarcheological series were subjected to paleomicrobiological analysis to establish the presence of MTBC bacteria. As all individuals showing clear osteological signs of TB infection belonged to a single grave group, 38 individuals from this grave group were analysed. The sample included the case of Pott's disease as well as individuals both with and without osseous TB manifestations. The detection of TB DNA in the individual with Pott's disease provided further evidence for the occurrence of TB in Neolithic populations of Europe. Moreover, our molecular analysis indicated that several other individuals of the same grave group were also infected with TB, opening the possibility for further analyses of this unique Neolithic skeletal series.

Langerhans cell histiocytosis or tuberculosis on a medieval child (Oppidum de la Granède, Millau, France - 10th-11th centuries AD).

In 2008, a skeleton of a 1 - 2.5-year-old child radiocarbon dated from the 10th - 11th century AD was discovered on the oppidum of La Granède (Millau, France). It presents multiple cranial osteolytic lesions having punched-out or geographical map-like aspects associated with sequestrum and costal osteitis. A multi 3D digital approach (CT, µCT and virtual reconstruction) enabled us to refine the description and identify the diploic origin of the lytic process. Furthermore, precise observation of the extent of the lesions and associated reorganization of the skeletal micro-structure were possible. From these convergent pieces of evidence, the differential diagnosis led to three possibilities: Langerhans cell histiocytosis, tuberculosis, or Langerhans cell histiocytosis and tuberculosis.

Three-dimensional imaging of past skeletal TB: From lesion to process.

3D imaging has become an essential tool in the field of biological anthropology, notably for human evolution purposes. High resolution virtual 3D reconstructions of original specimens contribute to their preservation and broaden the ability for research, teaching and exchanges. Paleopathology can get substantial benefit from these methods, among others for reconstructing infectious pathological processes on ancient bones. Tuberculosis is frequently diagnosed on ancient human remains; however, some osseous expressions are difficult to interpret using classical methods. We illustrate here the interest of 3D methods for reconstructing processes involved in pathological bone changes due to Mycobacterium tuberculosis infection. Four paleopathological specimens attributed to this infection, dating from different time periods and concerning diverse parts of the skeleton have been analyzed using a specific 3D digital chain we have previously developed. These 3D analyses allow to virtually reconstruct the initial location and aspect of the infectious process, its extension as well as its possible diffusion to the surrounding soft tissues. This possible virtual follow-up of the disease leads to the concept of processual paleopathology that we would like to introduce in the field. The 3D methodology can help to improve our knowledge of natural history and evolution of ancient human infections such as tuberculosis.