A Mediaeval Case of Lepromatous Leprosy from 13–14th Century Orkney, Scotland

Erosion in the 1960s resulted in exposure of human skeletal remains from a Norse Christian cemetery at Newark Bay, Orkney, Scotland. One set of remains showed osteological evidence of advanced lepromatous leprosy, but the absence of bones from the lower limbs precluded definitive diagnosis. The aim of the present study was to determine whether Mycobacterium leprae could be detected in bone extracts, as a means of confirming the diagnosis of leprosy. Bone samples were examined from the suspected leprosy case and from a second contemporary burial thought to be free of disease. DNA was amplified by polymerase chain reaction (PCR) using primers specific for a repetitive element (RLEP) characteristic of M. leprae. Additional PCR tests specific for Mycobacterium tuberculosis and for amelogenin (a human gene suitable for sex determination) were also applied to the samples. M. leprae DNA was detected only in the skull sample from the suspected leprosy case. The DNA sequence was identical to that found in present day isolates of M. leprae. Positive results were obtained only using a PCR reaction designed to amplify relatively short stretches of DNA (<175 bp), suggesting the microbial DNA had undergone extensive fragmentation. There was no evidence of M. tuberculosis DNA in bones from the leprosy suspect or control individual. The ability to recover ancient samples of DNA provides an opportunity to study long-term evolutionary changes that may affect the epidemiology of microbial pathogens.     

Paleopathological and Molecular Study on Two Cases of Ancient Childhood Leprosy from the Roman and Byzantine Empires

This study is based on the paleaopathology of leprosy on human skeletal remains and the detection of ancient Mycobacterium leprae DNA. Two cases of childhood leprosy were recognized. The first case was in a Roman necropolis at Martellona (Rome, Central Italy), dated to the 2nd to 3rd centuries ce . The skeleton of a child aged 4–5 years, from tomb 162, is the youngest individual in Italy from this time period, with the clear rhino‐maxillary syndrome and other bony changes indicative of leprosy. The second case from a burial at Kovuklukaya, in the Sinop region of Northern Turkey, was from the 8th to the 10th centuries, during the Byzantine era. The endocranium of a 4–5‐month‐old infant with new bone formation—an indication of chronic inflammation—was positive for M. leprae DNA. Infant and childhood leprosy is uncommon today, and there is a scarcity of information in the osteoarchaeological literature of leprosy in the past, especially in children. The significance of these cases is that it adds to an understanding of the history of the disease in the former Roman Empire. It is hoped that over time sufficient data can be obtained to understand the epidemiological dynamics and clinical evolution of leprosy from the ancient period until today. Copyright © 2012 John Wiley & Sons, Ltd.     

A case study of possible differential diagnoses of a medieval skeleton from Denmark: leprosy,ergotism, treponematosis,sarcoidosis or smallpox?

This paper uses macroscopic and radiological examinations to provide differential diagnoses of pathological lesions in the skeleton of a young woman, 20–25 years of age, which triggered the Danish palaeopathologist Vilhelm Møller‐Christensen's interest in leprosy. The skeleton was incomplete, but the majority of bones of the upper body, as well as the skull, were present. The pathological changes consisted of medullary and cortical lytic foci, periosteal reaction and enhanced cortical density. The lesions were most extensive on the left side, especially around the elbow, wrist and scapula. Treponematosis, leprosy, smallpox, ergotism, rheumatoid arthritis, tuberculosis and sarcoidosis are all reviewed with regard to bone and joint pathology and their likelihood of being the correct diagnosis. We concluded that the most plausible diagnosis is treponematosis, but neither sarcoidosis nor smallpox can be completely excluded. Copyright © 2007 John Wiley & Sons, Ltd.     

Periostitis and osteolysis in a medieval skeleton from South‐West Hungary: (Leprosy,treponematosis, tuberculosis or hypertrophic osteoarthropathy) A diagnostic challenge!

This paper uses macroscopic and radiological examinations to provide differential diagnoses of pathological lesions in the well‐preserved skeleton of a young male from the medieval site of Zalavár in South‐West Hungary. Macroscopic inspection of the skeleton revealed conspicuous thickening of the tibiae and fibulae with ‘tree bark’ appearance of the cortex. Periosteal proliferations are also found on the calcanea and on the posterior part of the femora. The metatarsals showed bony proliferation and bone dissolution. No alteration of the axial skeleton or the skull was noted. Radiographs showed thickening of the cortex of tubular bones due to a multilayered type of periosteal apposition. The likelihood of these symptoms being the result of melorheostosis, hypervitaminosis A, fluorosis, thyroid acropachy, endosteal hyperostosis, tuberculosis (TB), hypertrophic osteoarthropathy (HOA), treponematosis and leprosy is reviewed. None completely explains the entire spectrum of pathological lesions in the current individual, but the individual may have suffered from two co‐existent diseases. As a clinical entity, the changes in the metatarsals are compatible with leprosy, whereas the periosteal proliferations of the lower limbs point to a diagnosis of HOA as a secondary syndrome. Thus, a combination of leprosy and TB is suggested as a potential diagnosis. Although not definitive, our differential diagnosis was able to exclude a number of conditions producing periosteal apposition. Copyright © 2011 John Wiley & Sons, Ltd.     

Variable nucleotide tandem repeat (VNTR) typing of two palaeopathological cases of lepromatous leprosy from Mediaeval England

Using ancient DNA methods, we have examined in detail two archaeological cases of leprosy from Mediaeval England. The first was a child skeleton with rhino-maxillary changes typical of lepromatous leprosy (LL). The second case was the skeleton of a male adult who showed both typical rhino-maxillary changes and osteitis/periostitis on the leg and foot bones. Bone powder was sampled from both cases and DNA extracts were prepared. These were subjected to a series of polymerase chain reactions (PCRs) specific for regions on the Mycobacterium leprae genome. The repetitive element RLEP was used for confirmation of M. leprae DNA and then three polymorphic regions were successfully amplified and sequenced to determine the number of variable nucleotide tandem repeats (vntr) at these loci. These were the microsatellite regions ML2344 and ML2172 and the minisatellite region ML0058. Genotyping data from the strains preserved within the skeletal remains were compared with those obtained for a reference strain of M. leprae. Variation at these three loci was found between both burials and the reference strain, indicating that vntr typing of LL cases from the archaeological record is a useful way of confirming disease and an additional means of authenticating aDNA data. This demonstrates the feasibility of targeting multiple loci for phylogenetic studies of leprosy strains from archival sources.     

Metastatic carcinoma in a leper skeleton from a Medieval cemetery in Chichester England

Both leprosy and metastatic cancer are well documented in the literature on human skeletal palaeopathology. The manifestation of both pathological conditions in a single archaeological skeleton has not been reported. A case from a Medieval site in Chichester, England exhibits bone lesions and patterns of skeletal involvement indicative of both these diseases. Evidence of leprosy is largely restricted to the lower tibiae, fibulae and the bones of the feet. Fine destructive foci and reactive fibre bone associated with metastatic cancer are distributed in many of the remaining areas of the skeleton. There is minimal overlap in the areas of the skeleton involved in the two pathological processes.     

Leprosy in a skeleton from the 7th century necropolis of Vicenne‐Campochiaro (Molise,Italy)

Analysis of the skeleton from tomb 144 of the early medieval necropolis of Vicenne‐Campochiaro in Central Italy revealed several features indicative of leprosy. The skeleton belongs to a male estimated to be between 20 and 25 years of age at death. The distal halves of the 1st and 2nd left metatarsals present acro‐osteolysis and both legs show severe subperiosteal bone reaction. The facial skeleton shows changes compatible with a chronic inflammatory process, possibly due to an infectious disease. The anatomical distribution of the lesions and their association with other skeletal lesions seems to be compatible with a near‐lepromatous form of leprosy. A differential diagnosis is made, and the skeletal traits pathognomonic of leprosy are discussed. Copyright © 2005 John Wiley & Sons, Ltd.     

Leprosy at the Lazaretto on St Eustatius,Netherlands Antilles

Leprosaria established in the Americas during the Colonial period bear many similarities with those found in medieval Europe. They are comparable in terms of isolation, the objectification of leprosy sufferers and their association with religious charities. The Lazaretto on St Eustatius was operated from 1866 to 1923. The site was investigated to recover palaeopathological evidence of leprosy at a leprosarium in the Americas. Five burials were excavated; three individuals showed evidence of bone modifications consistent with those caused by leprosy, including aspects of ‘rhinomaxillary syndrome’ and the bilaterally symmetrical post‐cranial changes that have been described in leprosy examples from medieval Europe. An exceptional find was the presence of potentially leprous bone changes to the hyoid, thyroid and 3^rd–6^th cervical vertebrae. Copyright © 2007 John Wiley & Sons, Ltd.     

Earliest evidence of spinal tuberculosis from the Aneolithic Yayoi period in Japan

Tuberculosis has existed from early prehistoric days to modern times. The main causative agents of tuberculosis worldwide are Mycobacterium tuberculosis (M. tuberculosis) and M. bovis, along with M. africanum, M. cenettii and M. microti; these species make up the ‘M. tuberculosis complex’. This worldwide infection has been of special interest to palaeopathologists due to its characteristic bone lesions as well as its great antiquity. Historically, tuberculosis has been recognised in Japan for more than a thousand years. However, the origin and early prevalence of tuberculosis remain unknown. In the present study, we present the earliest evidence of skeletal tuberculosis found in the Aneolithic Yayoi period in Japan (ca. 300 BC to AD 300). The skeletal remains showing typical pathological changes of spinal tuberculosis were dated to between 454 BC and AD 124 by dendrochronological methods using coburied arrow-shield board and house columns made of Japanese cedar. We discuss the early prevalence of this infectious disease and its influence on the population history of the Japanese from prehistoric to Aneolithic times. Copyright © 2006 John Wiley & Sons, Ltd.     

Pre‐Columbian Tuberculosis in Northwest Argentina: Skeletal Evidence from Rincón Chico 21 Cemetery

Systematic excavation of collective burial sites makes possible the recovery of skeletal series which may show bony evidence of infectious pathological conditions. This paper presents the first evidence of the existence of tuberculosis in prehistoric populations of NW Argentina with a subsistence economy based on agriculture and pastoralism. The study was carried out on individuals from Rincón Chico 21 cemetery, a burial site located in the Santa María Valley, Catamarca, used between the Late Ceramic Period and the onset of the Inca empire expansion (AD 1000–1400). Six individuals out of the 70 so far excavated showed destructive lesions in the vertebral bodies and periosteal reactions in other bones. The morphology and distribution of bone lesions led us to rule out several diseases from a broad spectrum of possible diseases that could have affected the skeletal system. Thus, the lesions were interpreted as caused by mycobacterial infections (Mycobacterium tuberculosis Complex). Considering previous studies on the dynamics of biocultural interactions which take into account information related from contextual associations and chronology, we can conclude that a tuberculosis‐like disease was present in prehistoric populations from NW Argentina. Copyright © 2011 John Wiley & Sons, Ltd.     

Necropoli of Pill'e Matta Quartucciu (Cagliari,Sardinia): wild bee and solitary wasp activity and bone diagenetic factors

During the excavation at the necropolis and the laboratory studies, a variety of damage was found in the human bone remains. This damage included round holes, irregular bone destruction and irregular etching. This can be found especially in the skulls and in the long bones, and sometimes in the vertebrae and on the hands and feet. The exceptionality of this demanded meticulous study in order to define whether it was a diagenetic phenomenon of biological origin. Further studies revealed that the damage to the bones was produced by three kinds of Hymenoptera that build galleries: one of these belongs to the Sphecidae family, the other two to the Halictidae family. During the excavation of a grave it had been observed that a Halictidae had invaded the burial context and consequently contributed to the taphonomic and diagenetic process. The laboratory activities were directed towards the measurement of the damage to the human bone remains in 45 tombs. A histogram shows that the holes are not the result of accidental lesions, and the more frequent percentage values show similarities with the dimensions of holes built by the insects. The observations and the results of the study at the archaeological site enabled me to establish how significant the role of the insects had been, in particular of the Hymenoptera, in modifying the condition of the bone remains. Copyright © 2008 John Wiley & Sons, Ltd.     

Deficiencies and challenges in the study of ancient tuberculosis DNA

We explore the standards of research and reporting needed to justify the destructive analysis of archaeological human bone for biomolecular studies of ancient tuberculosis (TB). Acceptable standards in osteological interpretation have been met in some biomolecular papers, but there are also cases where insufficient care has been taken in distinguishing between pathognomonic lesions and those that are ‘consistent with’ a diagnosis of TB. Some biomolecular studies have failed to recognize that archaeological bones might be contaminated with environmental mycobacteria whose DNA could give rise to false positives in polymerase chain reactions directed at members of the Mycobacterium tuberculosis complex. The difficulties of applying spoligotyping to ancient DNA have also been underestimated and conclusions drawn from such analyses are often weakly supported. Assumptions that mycobacterial DNA preserves better than human DNA, and that contamination with modern DNA is less of a problem, has led in some cases to a laxity in research standards with insufficient attention paid to the need to authenticate ancient DNA results. We illustrate our concerns by reference to a recent paper reporting biomolecular detection of ancient TB DNA in skeletons from the eastern Mediterranean Neolithic settlement of Atlit-Yam. We are unconvinced that the skeletal evidence presented in this paper gives sufficient indication of TB to warrant destructive analysis, and we are concerned that during the biomolecular part of the project inadequate attention was paid to the possibility that results might be due to laboratory cross-contamination or to amplification of environmental mycobacterial DNA present in the bones.     

The effect of growth on stable nitrogen isotope ratios in subadult bone collagen

Stable nitrogen isotopes have been used to reconstruct infant feeding practices as nursing infants have elevated δ¹⁵N ratios compared with their mothers. However, infancy is also a time of rapid growth, which may alter nitrogen isotope diet‐to‐tissue spacing. Several studies have documented a decrease in δ¹⁵N during growth in tissues with relatively fast accretion rates. This study investigates the effect that the growth of long bones, via collagen accretion, has on δ¹⁵N ratios. Long bones from individuals aged seven to nineteen years were obtained from a protohistoric ossuary in Ontario, Canada. Analysis of juveniles and adolescents permitted the examination of growth in a group who were not also nursing. It is concluded that a nitrogen isotope growth effect is not detectable in bone collagen from juveniles and adolescents, because: (1) δ¹⁵N ratios are not significantly different among the epiphyses, metaphyses and diaphysis of a growing long bone; (2) δ¹⁵N ratios are not significantly different between faster‐growing versus slower‐growing metaphyses; and (3) δ¹⁵N ratios are not significantly different between bones (or areas of a bone) that are still undergoing growth, versus bones that have ceased growing. The relatively slow speed of collagen accretion may explain why a growth effect is not manifested. Ultimately this research lends support to the use of nitrogen isotopes from bone collagen for infant feeding reconstructions. Copyright © 2009 John Wiley & Sons, Ltd.     

Bone Modifications Resulting from Captive Chimpanzee Mastication: Implications for the Interpretation of Pliocene Archaeological Faunas

In order to obtain chimpanzee (Pan troglodytes) mastication damage on bones, cleaned, disarticulated ribs and long bones of bovids and cervids were coated with food substances found palatable by captive chimpanzees. The bones were then presented to four groups of mixed sex and mixed age chimpanzees from the Tulsa Zoo (Oklahoma) and the Southwest Foundation for Biomedical Research (San Antonio, Texas). In the process of feeding on the coated bones, the chimpanzees inflicted discernible mastication damage on 73·61% of the total recovered bone specimens. Analysis of these chimpanzee-induced bone modifications reveals that non-technological hominoids of the same approximate size and with comparable dentition and bite force to the gracile australopithecines and earliestHomoare very capable of inflicting the same range and degree of damage to bones as are feeding carnivores. This finding implies that zooarchaeologists must take a contextual (configurational) approach when analysing mastication damage on arch-aeological bones, rather than automatically attributing all such damage to carnivores.     

Preservation of bird bones: erosion versus digestion by owls

Surface damage to bird bones from pellets of three species of owls (S. aluco, A. otus and B. bubo) was compared with that of bird bones exposed to weathering and soil corrosion (Záskogo Cave in the Western Balkan Mountains, Bulgaria and Ciemna Cave, southern Poland). Scanning electron microscopy and light microscope examinations indicate that the effects of weathering of bird bones (extensive pitting and flaking away on whole bones, holes with sharp edges, depressions with rough bottoms and sharp edges of breakage) are distinguishable from those of digestion (rounding of hole edges on articular ends and sometimes on shafts, rounding of breakage). However, soil corrosion may produce similar rounding to that of digestion, which may hamper taphonomic interpretations of fossil assemblages. Two stages of weathering in bird bone are distinguished. Generally, the kind of damage done to bird bones is similar to that done to mammalian remains. Taphonomic studies of fossil bird remains should take into account a combination of features, including surface damage, fragmentation and possible chemical alterations of bone tissues. © 1997 John Wiley & Sons, Ltd.     

The Red Fox (Vulpes vulpes) as an Accumulator of Bones in Cave‐like Environments

Identifying the behavioural patterns of bone collecting animals is a crucial aspect of taphonomic studies. Although many studies have established criteria for identifying animal‐collected or animal‐modified bones, very few papers describe the distinguishing features of fox‐made bone assemblages. The bone assemblage collected in an inactive underground stone mine in Potok‐Senderki (Poland) is diagnostic of a red fox (Vulpes vulpes) den. This site provides an ideal opportunity to develop an understanding of the bone collecting behaviour of red foxes in cave‐like environments. This study showed that bones collected by red foxes are concentrated in clusters. The bones represent a broad spectrum of local fox prey species, with most bones showing the marks of gnawing. Each cluster may contain from <10 to >100 bones. Furthermore, the long axes of the bones in clusters frequently show specific orientation. The analysis of bones at this site might make an important contribution towards the establishment of baseline criteria for the identification and evaluation of fox‐accumulated bone assemblages. Copyright © 2012 John Wiley & Sons, Ltd.     

The limits of biomolecular palaeopathology: ancient DNA cannot be used to study venereal syphilis

To determine whether ancient DNA (aDNA) can be used to study the palaeopathology of venereal syphilis, we carried out a comprehensive analysis of the preservation of human and pathogen DNA in a set of 46 bones of various ages, most of which displayed osteological indications of the disease. Bones came from seven English cemetery sites that were in use during the 9th–19th centuries. Twelve of the 46 bones consistently yielded mitochondrial DNA (mtDNA) sequences after replicate polymerase chain reactions (PCRs), and a further 13 bones yielded mtDNA sequences with less reproducibility. The sequence data enabled tentative mitochondrial haplogroups to be assigned to nine of the bones, and the identities and frequencies of these haplogroups were compatible with the geographical origins of the bones. Twenty-one bones consistently gave negative results with all mtDNA PCRs, indicating that at least these bones were not contaminated with modern human DNA, and those bones that gave positive results only yielded one sequence each, again suggesting that widespread modern contamination had not occurred. Mycobacterium tuberculosis sequences were obtained from seven bones, including three of five bones with tuberculous lesions. The cloned and direct sequences obtained from both the mtDNA and M. tuberculosis PCR products showed features typical of degraded aDNA. All of these results suggest that at least some of the 46 bones that we studied were suitable for aDNA analysis. All 46 bones were tested with nine different treponemal PCRs, each optimised to give a detection limit of ≤5 genomes. Although various bones gave PCR products of the expected size with one or more of these PCRs, sequencing showed that none of these products were authentic treponemal amplicons. Our failure to detect treponemal DNA in bones that were suitable for aDNA analysis, using highly sensitive PCRs, suggests that treponemal DNA is not preserved in human bone and that it is therefore not possible to use aDNA analysis to study venereal syphilis. Any past or future paper claiming detection of treponemal aDNA should therefore be accompanied by a detailed justification of the results.     

THERMAL ALTERATIONS IN ARCHAEOLOGICAL BONES*

Archaeological bones, found close to Mexico City (Tlatelcomila), with alterations due to probable thermal exposure attributed to a possible case of cannibalism, have been characterized by XRD, SAXS and SEM. It is shown how these non‐destructive techniques, which are not conventionally used in archaeological research, may provide useful information. The deterioration degree of the bone structure is quantified through parameters such as gyration radius or fractal dimension. As some of the reported modifications could be attributed to diagenesis, a discussion is presented in terms of bone crystallinity and ionic exchange. Furthermore, the hydroxyapatite crystallite size increased depending on colour: this feature is not explained by diagenesis—it can only be attributed to temperature. We conclude that the samples were thermally treated. Indeed, thermal treatment alters the structure and morphology of bone at a very fine level (microscopic and nanometric), while morphology follows the structural modifications. It is determined that the studied human bones were treated at three different temperatures close to 100, 250 and 600°C. Therefore, they were either boiled (barbacoa) or grilled.