Ancient genomes reveal a high diversity of Mycobacterium leprae in medieval Europe.

Studying ancient DNA allows us to retrace the evolutionary history of human pathogens, such as Mycobacterium leprae, the main causative agent of leprosy. Leprosy is one of the oldest recorded and most stigmatizing diseases in human history. The disease was prevalent in Europe until the 16th century and is still endemic in many countries with over 200,000 new cases reported annually. Previous worldwide studies on modern and European medieval M. leprae genomes revealed that they cluster into several distinct branches of which two were present in medieval Northwestern Europe. In this study, we analyzed 10 new medieval M. leprae genomes including the so far oldest M. leprae genome from one of the earliest known cases of leprosy in the United Kingdom-a skeleton from the Great Chesterford cemetery with a calibrated age of 415-545 C.E. This dataset provides a genetic time transect of M. leprae diversity in Europe over the past 1500 years. We find M. leprae strains from four distinct branches to be present in the Early Medieval Period, and strains from three different branches were detected within a single cemetery from the High Medieval Period. Altogether these findings suggest a higher genetic diversity of M. leprae strains in medieval Europe at various time points than previously assumed. The resulting more complex picture of the past phylogeography of leprosy in Europe impacts current phylogeographical models of M. leprae dissemination. It suggests alternative models for the past spread of leprosy such as a wide spread prevalence of strains from different branches in Eurasia already in Antiquity or maybe even an origin in Western Eurasia. Furthermore, these results highlight how studying ancient M. leprae strains improves understanding the history of leprosy worldwide

Ancient genomes reveal a high diversity of Mycobacterium leprae in medieval Europe.

Studying ancient DNA allows us to retrace the evolutionary history of human pathogens, such as Mycobacterium leprae, the main causative agent of leprosy. Leprosy is one of the oldest recorded and most stigmatizing diseases in human history. The disease was prevalent in Europe until the 16th century and is still endemic in many countries with over 200,000 new cases reported annually. Previous worldwide studies on modern and European medieval M. leprae genomes revealed that they cluster into several distinct branches of which two were present in medieval Northwestern Europe. In this study, we analyzed 10 new medieval M. leprae genomes including the so far oldest M. leprae genome from one of the earliest known cases of leprosy in the United Kingdom-a skeleton from the Great Chesterford cemetery with a calibrated age of 415-545 C.E. This dataset provides a genetic time transect of M. leprae diversity in Europe over the past 1500 years. We find M. leprae strains from four distinct branches to be present in the Early Medieval Period, and strains from three different branches were detected within a single cemetery from the High Medieval Period. Altogether these findings suggest a higher genetic diversity of M. leprae strains in medieval Europe at various time points than previously assumed. The resulting more complex picture of the past phylogeography of leprosy in Europe impacts current phylogeographical models of M. leprae dissemination. It suggests alternative models for the past spread of leprosy such as a wide spread prevalence of strains from different branches in Eurasia already in Antiquity or maybe even an origin in Western Eurasia. Furthermore, these results highlight how studying ancient M. leprae strains improves understanding the history of leprosy worldwide

Case Reports: Legg-Calvé-Perthes Disease in Czech Archaeological Material

Legg-Calvé-Perthes disease (osteochondrosis of the femoral head) has been recognized in archaeological material for nearly a century but is extremely rare. We describe two Czech cases from archaeological findings. The first case was diagnosed in the skeleton of a man older than 50 years with the left hip affected. The skeleton was in grave Number 2 of the Langobard cemetery at Lužice (Moravia) and dated to the end of the fifth century and the beginning of the sixth century AD. The second case was described by J. Chochol in 1957 on the left femur and half of the pelvis of a skeleton from an archaeological investigation in Brandýsek (Bohemia), ninth to tenth centuries AD. Using the diagnostic criteria of Ortner and Putschar, we excluded slipped capital femoral epiphysis in both cases. We discuss the differential diagnosis of Legg-Calvé-Perthes disease versus unilateral and bilateral osteochondroses of the femoral head in archaeological and current clinical material

Ancient genomes reveal a high diversity of Mycobacterium leprae in medieval Europe.

Studying ancient DNA allows us to retrace the evolutionary history of human pathogens, such as Mycobacterium leprae, the main causative agent of leprosy. Leprosy is one of the oldest recorded and most stigmatizing diseases in human history. The disease was prevalent in Europe until the 16th century and is still endemic in many countries with over 200,000 new cases reported annually. Previous worldwide studies on modern and European medieval M. leprae genomes revealed that they cluster into several distinct branches of which two were present in medieval Northwestern Europe. In this study, we analyzed 10 new medieval M. leprae genomes including the so far oldest M. leprae genome from one of the earliest known cases of leprosy in the United Kingdom-a skeleton from the Great Chesterford cemetery with a calibrated age of 415-545 C.E. This dataset provides a genetic time transect of M. leprae diversity in Europe over the past 1500 years. We find M. leprae strains from four distinct branches to be present in the Early Medieval Period, and strains from three different branches were detected within a single cemetery from the High Medieval Period. Altogether these findings suggest a higher genetic diversity of M. leprae strains in medieval Europe at various time points than previously assumed. The resulting more complex picture of the past phylogeography of leprosy in Europe impacts current phylogeographical models of M. leprae dissemination. It suggests alternative models for the past spread of leprosy such as a wide spread prevalence of strains from different branches in Eurasia already in Antiquity or maybe even an origin in Western Eurasia. Furthermore, these results highlight how studying ancient M. leprae strains improves understanding the history of leprosy worldwide

A migration-driven model for the historical spread of leprosy in medieval Eastern and Central Europe

Leprosy was rare in Europe during the Roman period, yet its prevalence increased dramatically in medieval times. We examined human remains, with paleopathological lesions indicative of leprosy, dated to the 6th-11th century AD, from Central and Eastern Europe and Byzantine Anatolia. Analysis of ancient DNA and bacterial cell wall lipid biomarkers revealed Mycobacterium leprae in skeletal remains from 6th-8th century Northern Italy, 7th-11th century Hungary, 8th-9th century Austria, the Slavic Greater Moravian Empire of the 9th-10th century and 8th-10th century Byzantine samples from Northern Anatolia. These data were analyzed alongside findings published by others. M. leprae is an obligate human pathogen that has undergone an evolutionary bottleneck followed by clonal expansion. Therefore M. leprae genotypes and sub-genotypes give information about the human populations they have infected and their migration. Although data are limited, genotyping demonstrates that historical M. leprae from Byzantine Anatolia, Eastern and Central Europe resembles modern strains in Asia Minor rather than the recently characterized historical strains from North West Europe. The westward migration of peoples from Central Asia in the first millennium may have introduced different M. leprae strains into medieval Europe and certainly would have facilitated the spread of any existing leprosy. The subsequent decline of M. leprae in Europe may be due to increased host resistance. However, molecular evidence of historical leprosy and tuberculosis co-infections suggests that death from tuberculosis in leprosy patients was also a factor

Results of the genome-wide analysis for the samples with sufficient coverage.

<p>Results of the genome-wide analysis for the samples with sufficient coverage.</p

Time to the most recent common ancestor (tMRCAs) for the entire <i>M</i>. <i>leprae</i> tree and individual branches (HPD = Highest Posterior Density).

<p>Time to the most recent common ancestor (tMRCAs) for the entire <i>M</i>. <i>leprae</i> tree and individual branches (HPD = Highest Posterior Density).</p

Worldwide distribution of the ancient and modern <i>M</i>. <i>leprae</i> strains analyzed in this study.

<p>Skulls represent strains from osteological specimens dated to the Medieval Period. Human silhouettes represent modern strains, sized to scale according to the number of samples, ranging from 1 (e.g. India) to 36 (South America) Animal silhouettes represent strains from the red squirrel, the nine-banded armadillo, and naturally infected nonhuman primates (a chimpanzee from Sierra Leone, a sooty mangabey from West Africa, and a cynomolgus macaque from The Philippines). Skulls outlined in black are the new <i>M</i>. <i>leprae</i> genomes reconstructed in this study, while skulls outlined in blue represent previously sequenced ancient genomes. Grey skulls are leprosy samples from this study that did not yield sufficient sequence for whole-genome analysis. The main <i>M</i>. <i>leprae</i> lineages, represented by branches (see <a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1006997#ppat.1006997.g002" target="_blank">Fig 2</a>) are color-coded.</p

Phylogenetic analysis of ancient and modern <i>M</i>. <i>leprae</i> strains.

<p>(<b>A</b>) Maximum parsimony tree reconstructed from 3124 informative SNP positions. The tree is drawn to scale, with branch lengths representing number of substitutions. <i>M</i>. <i>lepromatosis</i> was used as outgroup. The novel strains from this study are labelled in red, and the previously published ancient strains are labelled in blue. Animal symbols indicate strains isolated from red squirrels, armadillos and non-human primates. The main branches are color-coded, and the SNP subtypes are marked with dotted vertical bars. Bootstrap values (500 replicates) are shown next to each node. (<b>B</b>) Bayesian phylogenetic tree based on 2371 SNP positions calculated with BEAST 1.8.1. Median divergence times in years B.C.E. and C.E. are shown on the main nodes (the 95% Highest Posterior Density ranges are given in square brackets). Tip labels for each sample show the name, the country of origin and the isolation date, or the radiocarbon dates. The novel strains from this study are labelled in red, and the previously published ancient strains are labelled in blue. Posterior probabilities for each node are shown in grey. The main branches are color-coded. The hypermutator strains 85054, Amami, S15, Br14-3, Br2016-15, Zensho-4, Zensho-5 and Zensho-9 (as described in [<a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1006997#ppat.1006997.ref014" target="_blank">14</a>]) were excluded from this analysis.</p