A 14th century CE Brucella melitensis genome and the recent expansion of the Western Mediterranean clade.

Brucellosis is a disease caused by the bacterium Brucella and typically transmitted through contact with infected ruminants. It is one of the most common chronic zoonotic diseases and of particular interest to public health agencies. Despite its well-known transmission history and characteristic symptoms, we lack a more complete understanding of the evolutionary history of its best-known species-Brucella melitensis. To address this knowledge gap we fortuitously found, sequenced and assembled a high-quality ancient B. melitensis draft genome from the kidney stone of a 14th-century Italian friar. The ancient strain contained fewer core genes than modern B. melitensis isolates, carried a complete complement of virulence genes, and did not contain any indication of significant antimicrobial resistances. The ancient B. melitensis genome fell as a basal sister lineage to a subgroup of B. melitensis strains within the Western Mediterranean phylogenetic group, with a short branch length indicative of its earlier sampling time, along with a similar gene content. By calibrating the molecular clock we suggest that the speciation event between B. melitensis and B. abortus is contemporaneous with the estimated time frame for the domestication of both sheep and goats. These results confirm the existence of the Western Mediterranean clade as a separate group in the 14th CE and suggest that its divergence was due to human and ruminant co-migration.

Correction: The paradox of HBV evolution as revealed from a 16th century mummy.

[This corrects the article DOI: 10.1371/journal.ppat.1006750.].

The paradox of HBV evolution as revealed from a 16th century mummy.

Hepatitis B virus (HBV) is a ubiquitous viral pathogen associated with large-scale morbidity and mortality in humans. However, there is considerable uncertainty over the time-scale of its origin and evolution. Initial shotgun data from a mid-16th century Italian child mummy, that was previously paleopathologically identified as having been infected with Variola virus (VARV, the agent of smallpox), showed no DNA reads for VARV yet did for hepatitis B virus (HBV). Previously, electron microscopy provided evidence for the presence of VARV in this sample, although similar analyses conducted here did not reveal any VARV particles. We attempted to enrich and sequence for both VARV and HBV DNA. Although we did not recover any reads identified as VARV, we were successful in reconstructing an HBV genome at 163.8X coverage. Strikingly, both the HBV sequence and that of the associated host mitochondrial DNA displayed a nearly identical cytosine deamination pattern near the termini of DNA fragments, characteristic of an ancient origin. In contrast, phylogenetic analyses revealed a close relationship between the putative ancient virus and contemporary HBV strains (of genotype D), at first suggesting contamination. In addressing this paradox we demonstrate that HBV evolution is characterized by a marked lack of temporal structure. This confounds attempts to use molecular clock-based methods to date the origin of this virus over the time-frame sampled so far, and means that phylogenetic measures alone cannot yet be used to determine HBV sequence authenticity. If genuine, this phylogenetic pattern indicates that the genotypes of HBV diversified long before the 16th century, and enables comparison of potential pathogenic similarities between modern and ancient HBV. These results have important implications for our understanding of the emergence and evolution of this common viral pathogen.

Gout in Duke Federico of Montefeltro (1422-1482): a new pearl of the Italian Renaissance.

The article examines the truthfulness of historical accounts claiming that Renaissance Duke Federico of Montefeltro (1422-1482) suffered from gout. By direct paleopathological assessment of the skeletal remains and by the philological investigation of historical and documental sources, primarily a 1461 handwritten letter by the Duke himself to his personal physician, a description of the symptoms and Renaissance therapy is offered and a final diagnosis of gout is formulated. The Duke's handwritten letter offers a rare testimony of ancient clinical self-diagnostics and Renaissance living-experience of gout. Moreover, the article also shows how an alliance between historical, documental and paleopathological methods can greatly increase the precision of retrospective diagnoses, thus helping to shed clearer light onto the antiquity and evolution of diseases.

Metastatic Prostate Carcinoma from Imperial Rome (1st to 2nd Centuries AD).

OBJECTIVE: This study describes and discusses a rare case of metastatic carcinoma that affected the skeleton of an adult male recovered in the necropolis of Casal Bertone in Rome (Italy). The necropolis, which dates back to the Imperial Age (1st to 2nd century AD), is located near some residential structures and a large place identified as a fullery (fullonica). METHODS: Anthropological and paleopathological studies of the skeletal remains were performed via careful macroscopic, microscopic, radiological (X-ray and CT scan), and histological investigation. RESULTS: The skeleton displayed mixed osteoclastic and osteoblastic lesions that mainly involved the axial bones, in particular the sternum, the ribs, the spine, and the scapular and pelvic girdles. The anatomical distribution and the destructive and proliferative nature of the lesions suggested diffuse metastases arising from a soft-tissue primary cancer. The age and sex of the individual, as well as radiographic and histological pictures, allowed diagnosis of an advanced prostate cancer with extensively diffused bone metastases. CONCLUSION: At present, this is the only case of prostate cancer from the Imperial Age recovered in Rome.

On the history of gout: paleopathological evidence from the Medici family of Florence.

OBJECTIVES: Throughout history, gout has been referred to as the "disease of the kings", and has been clearly associated with the lifestyle of the aristocratic social classes. According to the written sources, several members of the famous Medici family of Florence suffered from an arthritic disease that contemporary physicians called "gout". A paleopathological study carried out on the skeletal remains of some members of the family, exhumed from their tombs in the Church of San Lorenzo in Florence, offered a unique opportunity to directly investigate the evidence of the arthritic diseases affecting this elite group. METHODS: The skeletal remains of several members of the family were examined macroscopically and submitted to x-ray investigation. RESULTS: The results of the study allowed us to ascertain that the so-called "gout of the Medici" should be considered the clinical manifestation of three different joint conditions: diffuse idiopathic skeletal hyperostosis, rheumatoid arthritis and uratic gout. In particular, uric acid gout was diagnosed in the Grand Duke Ferdinand I (1549-1609). Recently, a new case of this disease was diagnosed in Anton Francesco Maria (1618-1659), a probable illegitimate member of the family. CONCLUSIONS: With this new case, uratic gout was observed in 2 out of 9 adult males, leading to suppose that the disease should have been a common health problem within the family. The aetiology of the disease has to be searched in environmental factors, since both historical and paleonutritional studies demonstrated that the diet of this aristocratic court was rich in meat and wine.

Trichuris trichiura in a post-Colonial Brazilian mummy.

Trichuris trichiura is a soil-transmitted helminth which is prevalent in warm, moist, tropical and subtropical regions of the world with poor sanitation. Heavy whipworm can result either in Trichuris dysenteric syndrome - especially in children - or in a chronic colitis. In heavy infections, worms can spread proximally and may cause ileitis. Here we provide first microscopic evidence for a T. trichiura adult worm embedded in the rectum of a post-Colonial Brazilian adult mummy. During Colonial and post-Colonial times, many European chroniclers described a parasitic disease named Maculo whose symptomatology coincides with heavy helminthiasis. Based on our findings and on comparison of ancient textual evidence with modern description of heavy whipworm, we feel confident in considering that the two syndromes are expressions of the same pathological condition.

A historical case of amelogenesis imperfecta: Giovanna of Austria, Grand Duchess of Tuscany (1547-1578).

The skeletal remains of Giovanna of Austria (1547-1578), daughter of the Emperor Ferdinand I of Habsburg (1503-1564) and first wife of the Grand Duke of Tuscany, Francesco I (1541-1587), exhumed from the Basilica of San Lorenzo in Florence, were submitted to paleopathological study. Examination of the dentition, which was in a good state of preservation, showed maxillary retrognathism, together with a caries lesion, moderate periodontal disease, malposition of the upper second premolars and tooth wear. Furthermore, several horizontal grooves were observed in both the buccal and the lingual crown surfaces of almost all teeth, especially the anterior ones. The orthopantomogram showed hypomineralized enamel and alveolar bone loss. Two third-molar teeth were investigated using micro-computed tomography (micro-CT) analysis, revealing highly irregular enamel caps with reduced average thickness. The observed features suggest a diagnosis of hypoplastic amelogenesis imperfecta, a developmental condition affecting enamel formation.

Diets, stress, and disease in the Etruscan society: Isotope analysis and infantile skeletal palaeopathology from Pontecagnano (Campania, southern Italy, 730-580 BCE).

Susceptibility to morbidity and mortality is increased in early life, yet proactive measures, such as breastfeeding and weaning practices, can be taken through specific investments from parents and wider society. The extent to which such biosocialcultural investment was achieved within 1st millennium BCE Etruscan society, of whom little written sources are available, is unkown. This research investigates life histories in non-adults and adults from Pontecagnano (southern Italy, 730-580 BCE) in order to track cross-sectional and longitudinal breastfeeding and weaning patterns and to characterize the diet more broadly. Stable carbon and nitrogen isotope analysis of incrementally-sampled deciduous and permanent dentine (n = 15), bulk bone collagen (n = 38), and tooth enamel bioapatite (n = 21) reveal the diet was largely based on C3 staple crops with marginal contributions of animal protein. Millet was found to play a role for maternal diet and trajectories of breastfeeding and feeding for some infants and children at the site. The combination of multiple isotope systems and tissues demonstrates exclusive breastfeeding was pursued until 0.6 years, followed by progressive introduction of proteanocius supplementary foods during weaning that lasted between approximately 0.7 and 2.6 years. The combination of biochemical data with macroscopic skeletal lesions of infantile metabolic diseases and physiological stress markers showed high δ15Ndentine in the months prior to death consistent with the isotopic pattern of opposing covariance.

Quinto Tiberio Angelerio and new measures for controlling plague in 16th-century Alghero, Sardinia.

Plague, a zoonotic disease caused by the bacterium Yersinia pestis, has been responsible for at least 3 pandemics. During 1582-1583, a plague outbreak devastated the seaport of Alghero in Sardinia. By analyzing contemporary medical texts and local documentation, we uncovered the pivotal role played by the Protomedicus of Alghero, Quinto Tiberio Angelerio (1532-1617), in controlling the epidemic. Angelerio imposed rules and antiepidemic measures new to the 16th-century sanitary system of Sardinia. Those measures undoubtedly spared the surrounding districts from the spread of the contagion. Angelerio seems to have been an extremely successful public health officer in the history of plague epidemics in Sardinia.

Francesco Maria Fiorentini (1603-1673): An Italian physician in 'The Iron Century'.

The aim of this paper is to shed light on the figure of Francesco Maria Fiorentini, a 17th-century physician from Lucca (Tuscany, Italy) and member of the Iatromechanical School, who distinguished himself for his role during the plague and the typhus epidemics that spread throughout Italy in the first half of that century. His work must be contextualized in a precise historical moment, which marked the gradual transition of Western medicine from the archaism of Galenic doctrine to that of the Iatromechanical School, when the foundations started to be laid for an experimental type of medicine that based its assumptions on the direct observation of phenomena concerning the human body. In this work, we mainly focus on the medical biography of Fiorentini and on the reasons why he enjoyed great social prestige among the most prominent figures of his time. However, Fiorentini should also be remembered as a multifaceted scholar, as evidenced by his numerous writings, which underline his erudition in disparate fields of knowledge.

Solitary osteochondromas in paleo-oncology: A case report from 4th-century BCE Pontecagnano (southern Italy).

OBJECTIVE: This study presents a new case of solitary osteochondroma (SOC) identified on the proximal tibia of a 4th-century BCE individual from Pontecagnano (Salerno, Italy) with an aim to contribute to differential diagnosis of bone tumors in archeological contexts. MATERIALS: Paleopathological assessment of a male individual with an estimated age-at death of 45.9-62.9 years brought to light during the archaeological excavations in the funerary sector of 'Sica de Concillis' within the necropolis of Pontecagnano. METHODS: Macroscopic and radiographic analyses were performed for diagnostic purposes. RESULTS: The proximal portion of the right tibia exhibited a large exophytic bone formation extending from the anteromedial to the posteromedial portion of the diaphysis. The x-ray confirmed the lesion being characterized by regular trabecular bone tissue with cortico-medullary continuity. CONCLUSIONS: The lesion observed is diagnostic of sessile SOC, a neoplasm that must have had aesthetic and, possibly, neurovascular complications due to its large size. SIGNIFICANCE: By offering a detailed description of a case of tibial osteochondroma and discussing the possible complications the individual might have experienced during life, this study highlights the role and importance of benign bone tumors in paleo-oncology. LIMITATIONS: Histological analysis was not carried out in order to preserve the integrity of the affected tibia. SUGGESTIONS FOR FURTHER RESEARCH: More attention should be devoted to benign tumors in paleopathology as their occurrence and manifestations in the past will lead to a better understanding of their impacts on the quality of life of affected individuals and to greater knowledge of their natural history.

Infantile scurvy as a consequence of agricultural intensification in the 1st millennium BCE Etruria Campana.

The 1st millennium BCE in Italy was a time of agricultural intensification of staple cereal production which shaped sociocultural, political, and economic spheres of pre-Roman groups. The lifeways and foodways of the Etruscans, the greatest civilization in western Europe before Roman hegemony, are traditionally inferred from secondary written sources, funerary archaeology, archaeobotany, and zooarchaeology. However, no direct data extrapolated from the study of human skeletal remains are available to evaluate the extent to which agricultural intensification and decreased dietary diversity impacted health and the expression of skeletal indicators of metabolic disease. Macroscopic and radiological analyses were conducted on an archaeological skeletal sample of non-adults (n = 29) recovered from Pontecagnano (southern Italy) dating to the Orientalizing period (730-580 BCE). This allowed us to identify five cases of scorbutic non-adults and to assign diagnostic values to skeletal lesions of scurvy that have not been previously described in the literature. The onset of scurvy in the examined sample is related to the increased reliance of Etruscans on crops lacking vitamin C in this period of agricultural intensification. The skeletal expression of scurvy varied among the non-adults, with differences in location and disease severity; these were interpreted considering the age-at-death of the individuals coupled with feeding behaviors and interindividual variability.

A case of secondary hypertrophic osteoarthropathy from medieval Tuscany (central Italy, 10th-12th centuries CE).

OBJECTIVES: This study aims to provide a detailed evaluation of a case of secondary hypertrophic osteoarthropathy (HOA) and to explore insights into the presence and consequences of disease in medieval rural Italy. MATERIALS: The skeleton of a male (US 4405) with an estimated age at death of 51-69 years excavated from the medieval rural site of Pieve di Pava (Siena, Italy). METHODS: Macroscopic and radiological (x-ray, CT) analyses were performed. RESULTS: Symmetrical extensive periosteal new bone formation on the diaphyseal and metaphyseal regions of this individual's long bones; the lower limbs were more extensively and severely affected than the upper limbs and the distal segments were more severely altered in comparison to the proximal ones. CONCLUSIONS: The macroscopic and radiological features are highly consistent with a diagnosis of secondary HOA. SIGNIFICANCE: The excellent state of preservation allowed the evaluation of rarely noted skeletal manifestations of HOA and provided insight into aspects of rural life in medieval Italy. LIMITATIONS: Molecular analysis was not successful in sequencing the aDNA of tuberculosis, therefore the underlying primary cause of secondary HOA, whether pulmonary or extrapulmonary, remains obscure in this case. SUGGESTION FOR THE FUTURE RESEARCH: It is advisable to regularly revisit the data available from osteoarchaeological collections in order to identify further cases of HOA, along with to further investigate the known cases to search for the underlying primary disease.

Contextualizing bilateral asymmetry and gender: A multivariate approach to femoral cross-sectional geometry at rural Medieval Pieve di Pava, Italy.

OBJECTIVES: Our objective was to identify the relationship between biocultural factors of sex-gender and age and patterns of femoral cross-sectional geometry with historical evidence about labor and activity from an archeological skeletal sample excavated from the rural Medieval site Pieve di Pava. MATERIALS AND METHODS: The study site, Pieve di Pava, was a rural parish cemetery in Tuscany with osteoarcheological remains from the 7th to 12th centuries. Cross-sectional geometric analysis of femora from 110 individuals dated to the 10th-12th centuries were used to examine trends in bone quantity, shape, and bending strength between age and sex groups, as well as in clusters identified through Hierarchical Cluster Analysis (HCA). RESULTS: Overall, our study sample showed remarkable heterogeneity and our cluster analysis revealed a complex underlying structure, indicating that divisions of labor did not follow a strict gender binary in our sample. We found high levels of bilateral asymmetry in our sample in multiple cross-sectional areas for a significant proportion of the population. We found minimal differences between age groups or sex. DISCUSSION: Our results suggest that males and females had varied experiences of labor and work during their lives that did not reflect the strict binary gender roles sometimes documented for medieval Europe. One important axis of difference is the direction and magnitude of bilateral asymmetry observed in our femur sample, which is associated with divergent trends in section moduli and bone area measures.