Metagenomic and paleopathological analyses of a historic documented collection explore ancient dental calculus as a diagnostic tool.

Dental calculus is a microbial biofilm that contains biomolecules from oral commensals and pathogens, including those potentially related to cause of death (CoD). To assess the utility of calculus as a diagnostically informative substrate, in conjunction with paleopathological analysis, calculus samples from 39 individuals in the Smithsonian Institution's Robert J. Terry Collection with CoDs of either syphilis or tuberculosis were assessed via shotgun metagenomic sequencing for the presence of Treponema pallidum subsp. pallidum and Mycobacterium tuberculosis complex (MTBC) DNA. Paleopathological analysis revealed that frequencies of skeletal lesions associated with these diseases were partially inconsistent with diagnostic criteria. Although recovery of T. p. pallidum DNA from individuals with a syphilis CoD was elusive, MTBC DNA was identified in at least one individual with a tuberculosis CoD. The authenticity of MTBC DNA was confirmed using targeted quantitative PCR assays, MTBC genome enrichment, and in silico bioinformatic analyses; however, the lineage of the MTBC strain present could not be determined. Overall, our study highlights the utility of dental calculus for molecular detection of tuberculosis in the archaeological record and underscores the effect of museum preparation techniques and extensive handling on pathogen DNA preservation in skeletal collections.

Chronic Maxillary Sinusitis: A comparison of osteological and CT methods of diagnosis.

OBJECTIVE: To assess the differences between endoscopic and radiological methods of analysis for diagnosing chronic maxillary sinusitis (CMS) in archaeological skeletal remains. MATERIALS: 32 crania from a Dutch post-medieval rural population. METHODS: We assessed the presence of bone changes indicative of CMS (i.e., bone growth and bone resorption) both endoscopically and through computed tomography (CT), and then compared results. RESULTS: We observed moderate agreement between bone growth scores obtained through endoscopy and CT, and fair agreement when assessing bone resorption. CONCLUSIONS: CMS prevalence rates observed through CT may be comparable to rates assessed endoscopically, although caution is needed when making direct comparisons. SIGNIFICANCE: This is the first study comparing data obtained through endoscopy and radiological methods in the study of CMS, informing paleopathologists about potential biases in data comparison. LIMITATIONS: Our small sample size likely impacted results. SUGGESTIONS FOR FURTHER RESEARCH: Further research is advised to fully explore the comparability of endoscopic and radiological method of analysis in the study of sinusitis. DATA AVAILABILITY: The datasets generated and analyzed during the current study are available upon request.

Two cases of smallpox from 1540 CE circum-contact (early colonial) Northern Coastal Peru.

OBJECTIVE: This project seeks to create a differential diagnosis for lesions found on the skeletal remains of two children as a means to explore the presence of viral disease in 16th- century Peru. MATERIALS: Extremely well-preserved human remains of two children who died between the ages of 1-2 years old, recovered from the circum-contact (∼1540 CE) cemetery in Huanchaco, Peru. METHODS: Macroscopic and radiographic analysis. RESULTS: Both individuals present with cortical thickening, symmetrical destructive lesions, metaphyseal expansion, perforations, exposure of the medullary cavity, resorption of metaphyseal ends and necrosis of the long bones, and deposited reactive new bone. These features are consistent with osteomyelitis variolosa and bacterial osteomyelitis. CONCLUSIONS: Three features of Individuals IG-124 and IG-493 suggest a highly consistent diagnosis of osteomyelitis variolosa: multiple skeletal lesions, the historical context of the area, and the high mortality rate of non-adults in the circum-contact cemetery. SIGNIFICANCE: Although viral infections are ubiquitous and well documented historically, their etiologies are often difficult to determine in archaeological populations. Orthopoxvirus variola (smallpox) is one of the many viruses whose archaeological impact is still under explored in skeletal remains. LIMITATIONS: The absence of smallpox in other children from the Huanchaco cemetery creates difficulty in ascertaining true prevalence rates or information on potential outbreaks. SUGGESTIONS FOR FURTHER RESEARCH: Further research analyzing aDNA from calculus and/or residues using a DIP-GC-MS method might create a better understanding of how smallpox spread through the region.

Periodontitis and alveolar resorption in human skeletal remains: The relationship between quantitative alveolar bone loss, occlusal wear, antemortem tooth loss, dental calculus and age at death in a low socioeconomic status, modern forensic human collection from Yucatan.

OBJECTIVE: This paper assesses the relationship between the distance between the cemento-enamel junction and alveolar crest and risk factors commonly associated with periodontitis. MATERIALS: Eighty individuals between 28 and 92 years old with known biological sex and age were analyzed from a 20th century forensic human collection from Merida, Yucatan (Mexico). METHODS: Macroscopic assessment, along with metric analysis, was employed using a probe. RESULTS: Ante-mortem tooth loss was positively correlated with the distance between the cemento-enamel junction and alveolar crest, as was the presence of root calculus in females. CONCLUSIONS: Cemento-enamel junction to alveolar crest distance is not a reliable indicator of periodontitis since it is not directly related to periodontitis-causing infectious pathogens, and since ante-mortem tooth loss can affect root exposure. SIGNIFICANCE: This study demonstrates that a purely quantitative approach to diagnosing periodontitis in archaeological and forensic human remains can be misleading. LIMITATIONS: The skeletal collection is only representative of the low socioeconomic class of Merida, and its female cohort is underrepresented. In addition, because the Xoclan collection is modern, limitations (particularly with respect to tooth wear) of the applicability of these interpretations to older archaeological remains exist. SUGGESTION FOR FURTHER RESEARCH: A combination of quantitative and qualitative characteristics of alveolar bone is needed to reliably diagnose periodontitis in skeletal populations.

Porotic hyperostosis and cribra orbitalia in samples from the Late Hillfort Period sites at Detkovice - Za zahradama (district Prostejov) and Vídenská street (district Brno), Czech Republic.

Porotic hyperostosis and cribra orbitalia are pathological changes occurring on the human skull. These changes were observed and evaluated on skeletal remains from Detkovice - Za zahradama and Vídenská Street in Brno; both sites are dated back to the 10th to 12th centuries AD. A total of 605 subjects were assessed for age, sex, and the above-stated pathologies using standard methods. The influence of age and sex on the occurrence of these pathologies was examined statistically. Results indicated that at the site of Detkovice - Za zahradama, porotic hyperostosis, and cribra orbitalia do not depend on sex or age. However, at Vídenská Street in Brno, these pathologies do not depend on sex, but they depend on age so a higher incidence of pathologies in juveniles is observed. Differences between both sites could be caused by different numbers of evaluated individuals or different state of preservation of skeletal remains. The aetiology of the origin of these pathologies could not be determined by the methodology used here, but with the prevailing lower age of pathological subjects, a lack of nutrition with consequent absence of iron and developing anaemia might be the cause. Based on our statistical data, we can observe that the pathologies we studied occur more frequently in children older than newborns and younger infants. This may indicate that these studied pathologies arise only during the lifetime of the individual and do not have a prenatal occurrence.

Probable cases of tuberculosis in Early Medieval pastoralists of Eastern Europe.

Two anthropological collections of the 8-9th century AD from the forest-steppe area of the southern Eastern Europe has been the subject of analysis of the osteo-articular alterations concerning tuberculosis related lesions. According to archaeological data, the Mayaki and Dmitrievka groups are from the territory of Khazar Khaganate, which is well-known by historical data. It was a traditional population with settled pastoral farming. There were studied 292 adult human remains by macroscopic morphological and radiological methods. The completeness of the skeletons could have been better due to the intentional selection of their parts for the museum funds, so there were limitations for the differential diagnosis of TB by morphological criteria. In general, 31 individuals (20 men, 11 women) were marked as possible candidates with skeletal TB. The number of skeletons with possible specific infections from the two series is comparable, but according to sex distribution, there are some significant differences. If in the Dmitrievka group, there was approximately the same distribution of the number of cases of possible infected men and women; in the Mayaki group then there was a significant difference in the prevalence of the male sample. Various reasons can explain the observed differences. Firstly, we cannot rule out an error in the diagnosis of tuberculosis only by morphological methods since the preservation of the skeletons leaves much to be desired. Secondly, as it is known, only a few percent of tuberculosis patients show skeletal alterations, so the direct quantitative comparison cannot be adequate in paleopathological studies. The ambiguity of morphological criteria, particularly for destructive lesions of bone other than vertebrae, does not allow asserting that all identified cases result from TB. However, the results of differential diagnosis can underline that at least half of the skeletons have skeletal alterations most likely related to TB. The final analysis of the remains by aDNA methods will permit more certain confirmation of the diagnosis.

More than one millennium (2nd-16th century CE) of the White Plague in the Carpathian Basin - New cases, expanding knowledge.

The causative agent of tuberculosis is still a widespread pathogen, which caused the death of ca. 1.6 million people globally in 2021. The paleopathological study of human remains revealed the antiquity of the disease and its continuous presence throughout the history of humankind. The Carpathian Basin has always been a biocultural melting pot, since it has seen several migrations over the centuries, and served as a location of admixture and interaction for numerous populations of different cultures. Thus, this geographical territory is ideal for the examination of the coevolutionary processes of hosts and their pathogens. We aimed to reveal the spatial and temporal distribution of tuberculosis cases excavated inside the borders of Hungary between the 2nd and 16th centuries CE. We established a comprehensive database by collecting 114 already published cases and introducing 39 new cases. The involved cases include those that have been confirmed by different molecular methods, as well as possible infections that were identified based on the presence of macromorphological and radiological alterations. The progress of future molecular and paleopathological studies can be facilitated by our dataset, as it presents spatial and temporal information concerning the spread of the disease in the Carpathian Basin, as well as the biological profile and detailed paleopathological description of lesions illustrated by photo- and radiographs.

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].

The palaeopathology of industry, a perspective from Britain.

OBJECTIVES: This article considers the position of palaeopathology of ca. 1750AD onward within the subdiscipline of Industrial Archaeology, and reflects upon the relationship between skeletal palaeopathology and textual sources on disease prevalences. METHODS: It draws upon the author's experience in engaging with threat-led archaeology. It synthesises key elements of palaeopathological literature, emphasising contributions to the IJPP VSI 'Changes in Health with the Rise of Industry', and also the broader literature regarding Industrial Archaeology. RESULTS: Industrial Archaeology has seen a recent refocus to include not only a concentration upon technological aspects of industry but also increased emphasis the social context of industrialisation. This movement toward a placement of people as well as machines centre stage has resulted in an environment conducive for paleopathology to make a greater impact upon studies of the period. CONCLUSIONS: Palaeopathologists need to ensure that their biocultural work is orientated toward research goals of broader relevance if the impact of their work is to be maximised. We cannot directly align prevalence data generated from skeletal and and written sources; roles played by these two sources of evidence will depend, inter alia, upon the problems being investigated. SIGNIFICANCE: The success of 'Industrial Palaeopathology' will be measured by the extent to which human remains studies move toward centre stage within the broader discipline of Industrial Archaeology. LIMITATIONS: Multiple perspectives on disciplinary development are possible. Academic traditions, relationships between university- and threat led-sectors, and the opportunities and challenges engendered by working with human remains, differ in different countries.

Observer agreement on the morphology of porous cranial lesions: Results from a workshop at the 2019 meeting of the Paleopathology Association.

OBJECTIVE: This paper presents the results of a 2019 Paleopathology Association workshop that tested observer agreement on porous cranial lesion morphology and presence using multiple sets of existing guidelines for data collection. MATERIALS: Sixteen conference attendees of varying osteological experience served as observers. Three crania were assigned to each of four published guidelines for identifying and categorizing lesion morphology, for a total of twelve well-preserved human crania from the National Museum of Natural History Biological Anthropology Collections. METHODS: Observers assessed each cranium macroscopically according to its assigned set of guidelines. RESULTS: Observer concordance was higher using scoring guidelines with higher-quality photographs, such as the 2019 guidelines from Rinaldo and colleagues. CONCLUSIONS: Data collection guidelines with high-quality color photos may support greater reliability of researcher-generated data on macroscopic skeletal features. SIGNIFICANCE: The conclusions of any research study are only as reliable as the data on which they are based. This work highlights the need for ongoing practices of quality control in a field in which much data results from individual judgement calls. LIMITATIONS: Observer concordance is not a measure of observer accuracy. Sample size is insufficient to draw broadly generalizable conclusions on the reliability of data collected using the guidelines tested, and conference environments are not a facsimile of research settings. SUGGESTIONS FOR FURTHER RESEARCH: Iterative testing of methodological consistency using larger sample sizes and more non-pathological crania is advised to identify the factors that influence observer discordance and to improve guidelines for qualitative assessments.

Deficiency diseases in the Kyiv Rus' subadult population: The issue of the small sample effects.

This paper underlines the importance of the small sample effects consideration in paleopathological research providing an example of children from Kyiv Rus' cemeteries dated to the 10th-13th century AD. The study presents data introduction and systematization, i.e., construction of the statistically significant disease profiles combining the analyzed sites according to the distribution of anemia, scurvy, and rickets. Then we present usually provided (often 'associative') interpretations for the obtained results and question their reliability from the perspective of the small samples effects. This leads to the discussion of explanatory limits of bioarcheological research basing on the small sample analysis.

Historical microbiology: researching past bioevents by integrating scholarship (re)sources with paleomicrobiology assets.

The analysis of past epidemics and pandemics, either spontaneous or of human origin, may revise the physical history of microbiota and create a temporal context in our understanding regarding pathogen attributes like virulence, evolution, transmission and disease dynamics. The data of high-tech scientific methods seem reliable, but their interpretation may still be biased when tackling events of the distant past. Such endeavors should be adjusted to other cognitive resources including historical accounts reporting the events of interest and references in alien medical cultures and terminologies; the latter may contextualize them differently from current practices. Thus 'historical microbiology' emerges. Validating such resources requires utmost care, as these may be susceptible to different biases regarding the interpretation of facts and phenomena; biases partly due to methodological limitations.

Bacteria and viruses have always impacted humankind. They do this directly by causing illness or indirectly by destroying crops and threatening livestock. We can learn a lot by studying disease events of the past ­ for example, we can see how bacteria and viruses have changed over time and predict how they might change in the future. This knowledge could be important to understanding present disease events and predicting future ones. In this review, we propose the concept of 'historical microbiology', which encourages collaboration between scientists, doctors, historians and linguists to provide historical, linguistic and cultural context to our scientific understanding of diseases of the past.

Differential diagnosis for two 'holes in the head' of a child from 982 to 904 BP in northern South America.

OBJECTIVE: To present paleopathological evidence of a congenital anomaly with photographic support and a review that will help scholars to diagnose the condition. MATERIALS: Well-preserved skeletal remains of a child from central Colombia, dated 968-1046 CE. METHODS: Macroscopic examination and computerized axial tomography. RESULTS: Two holes were observed in the skull. CONCLUSIONS: The pathology is consistent with a neural tube defect or an inclusion cyst. SIGNIFICANCE: Neural tube defects and inclusion cysts, in paleopathology, are rarely reported in children. The preservation and origin of the individual make this case valuable. The photographic support and the review is useful for other scholars in the field. LIMITATIONS: It was not possible to determine a single cause. SUGGESTIONS FOR FUTURE RESEARCH: Review cases in identified osteological collections.

Deciphering Diets and Lifestyles of Prehistoric Humans through Paleoparasitology: A Review.

Parasites have affected and coevolved with humans and animals throughout history. Evidence of ancient parasitic infections, particularly, reside in archeological remains originating from different sources dating to various periods of times. The study of ancient parasites preserved in archaeological remains is known as paleoparasitology, and it initially intended to interpret migration, evolution, and dispersion patterns of ancient parasites, along with their hosts. Recently, paleoparasitology has been used to better understand dietary habits and lifestyles of ancient human societies. Paleoparasitology is increasingly being recognized as an interdisciplinary field within paleopathology that integrates areas such as palynology, archaeobotany, and zooarchaeology. Paleoparasitology also incorporates techniques such as microscopy, immunoassays, PCR, targeted sequencing, and more recently, high-throughput sequencing or shotgun metagenomics to understand ancient parasitic infections and thus interpret migration and evolution patterns, as well as dietary habits and lifestyles. The present review covers the original theories developed in the field of paleoparasitology, as well as the biology of some parasites identified in pre-Columbian cultures. Conclusions, as well as assumptions made during the discovery of the parasites in ancient samples, and how their identification may aid in better understanding part of human history, ancient diet, and lifestyles are discussed.

A rare case of calvarial tuberculosis from the Avar Age (8th century CE) cemetery of Kaba-Bitózug (Hajdú-Bihar county, Hungary) - Pathogenesis and differential diagnostic aspects.

The aim of our paper is to present and discuss in detail the bony changes indicative of tuberculosis (TB) that were identified in a skeleton (KB67), unearthed from grave 67 of the 8th-century-CE cemetery of Kaba-Bitózug (Hungary). Furthermore, to provide the differential diagnoses of the observed alterations, with special attention to the cranial osteolytic lesions. During the macro- and micromorphological examinations of KB67, the skull revealed three small, well-circumscribed, punched-out osteolytic lesions accompanied by endocranial granular impressions, abnormal blood vessel impressions, periosteal appositions, and cortical erosion. The postcranial skeleton exhibited osteolytic lesions, cortical remodelling and erosion, and signs of hypervascularisation in the spine. Based on the differential diagnosis of the cranial osteolytic lesions and their co-occurrence with endocranial and vertebral bony changes indicative of TB, they most likely resulted from tuberculous involvement of the frontal and left parietal bones. The morphologically established diagnosis was confirmed by a PCR analysis that provided evidence for the presence of Mycobacterium tuberculosis DNA in KB67. KB67, the first reported archaeological case with calvarial TB from the present-day territory of Hungary, gives us a unique insight into the occurrence of a rare manifestation of TB in the Avar Age of the Great Plain.

Demographics and significance of porotic hyperostosis as assessed by surface microscopy.

BACKGROUND: Examination of parietal surface anatomy has been limited because standard techniques have insufficient resolution to identify and characterize the structures of interest. Perspectives derived thereof have not clarified their nature. Surface microscopy is pursued as a nondestructive technique to assess the character and implications of porotic pores (referred to as porotic hyperostosis), which have been subject of much speculation. METHODS: The external surface of the skulls, selected on the basis of age and gender, from the Hamann-Todd human collection are examined by epi-illumination microscopy for surface pores and to assess correlation with age, ethnicity, gender, anemia, infection, cancer, hypertrophic bone disorders, renal disease, and fractures. RESULTS: Pore-like surface defects are present in 2.7%-5% of individuals in the third-fifth decades of life; 7%, in the sixth-eighth; and 25%, in the ninth-11th, but absent in the second decade of life. They are gender and birthdate independent, but slightly more common in African Americans. Fractures are more common among individuals with parietal pores, while tuberculosis, cancer, and hypertrophic bone diseases and anemia are less common. DISCUSSION: This is the first study to actually examine the prevalence of parietal pores as a function of known age, race, and sex and provides a baseline for comparison with populations in which those variables are not clearly identifiable. While some porotic pores may be related to marrow hyperplasia, transcortical circulation may explain the majority.

Verification of tuberculosis infection among Vác mummies (18th century CE, Hungary) based on lipid biomarker profiling with a new HPLC-HESI-MS approach.

Tuberculosis (TB) was a large burden of infections that peaked during the 19th century in Europe. Mummies from the 18th century CE, discovered in the crypt of a church at Vác, Hungary, had high TB prevalence, as revealed by amplification of key fragments of TB DNA and genome-wide TB analysis. Complementary methods are needed to confirm these diagnoses and one approach uses the identification of specific lipid biomarkers, such as TB mycocerosic acids (MCs). Previously, MC derivatives were profiled by specialised gas chromatography-mass spectrometry (GC-MS), so an alternative more direct approach has been developed. Underivatized MCs are extracted and analysed by high-performance liquid chromatography linked to a mass spectrometer, in heated electrospray ionisation mode (HPLC-HESI-MS). The method was validated using representatives of the Mycobacterium tuberculosis complex and other mycobacteria and tested on six Vác mummy cases, previously considered positive for TB infection. Analysing both rib and soft tissue samples, four out of six cases gave profiles of main C32 and major C29 and C39 mycocerosates correlating well with those of M. tuberculosis. Multidisciplinary methods are needed in the diagnosis of ancient tuberculosis; this new protocol accesses important confirmatory evidence, as demonstrated by the confirmation of TB in the Vác mummies.

A dual process model for paleopathological diagnosis.

OBJECTIVES: This paper aims to provide an explicit theoretical model for the cognitive processes involved in paleopathological diagnosis. METHODS: The approach adopted is a dual process model (DPM). DPMs recognize that cognition is a result of both Type 1 (intuitive) and Type 2 (analytical) processes. DPMs have been influential for understanding decision-making in a range of fields, including diagnosis in clinical medicine. Analogies are drawn between diagnosis in a clinical and a paleopathological setting. RESULTS: In clinical medicine, both Type 1 and Type 2 processes play a part in diagnosis. In paleopathology the role of Type 1 processes has been unacknowledged. However, like clinical diagnosis, paleopathological diagnosis is inherently a result of a combination of both Type 1 and Type 2 processes. A model is presented by which Type 1 processes can be explicitly incorporated into a scientific approach to diagnosis from skeletal remains, and in which diagnosis is formalized as a process of hypothesis testing. SIGNIFICANCE: Accurately modelling our diagnostic processes allows us to understand the biases and limitations in our work and potentially helps us to improve our procedures, including how we impart diagnostic skills in pedagogical settings. LIMITATIONS: This work provides a theoretical model for paleopathological diagnosis. However, such models are by their nature dynamic and developing rather than static entities; it is hoped that this work stimulates further debate and discussion in this important area.

Tracking down the White Plague. Chapter two: The role of endocranial abnormal blood vessel impressions and periosteal appositions in the paleopathological diagnosis of tuberculous meningitis.

Although endocranial abnormal blood vessel impressions (ABVIs) and periosteal appositions (PAs) have been considered as paleopathological diagnostic criteria for tuberculous meningitis (TBM) based on findings of previous studies, they are not pathognomonic for tuberculosis (TB). Therefore, their utilization in the paleopathological practice can be questioned, especially in consideration that most of the previous studies were not performed on identified skeletal collections but on osteoarchaeological material and did not include statistical data analysis. To fill the aforementioned research gap, for the first time, a macroscopic investigation was conducted on identified pre-antibiotic era skeletons from the Terry Collection. A sample set of 234 individuals who died of TB (TB group) and 193 individuals who died of non-tuberculous causes (NTB group) were examined. The frequency of ABVIs and PAs, as well as other probable TB-related lesions was recorded. To determine the significance of difference (if any) in the frequencies of ABVIs and PAs between the two groups, χ2 testing of our data was performed. We found that ABVIs, PAs, and their co-occurrence with each other and with other probable TB-related lesions were more common in the TB group than in the NTB group. In addition, the χ2 comparative frequencies of ABVIs and PAs revealed a statistically significant difference between individuals who died of TB and individuals who died of NTB causes. Our findings strengthen those of previous studies that ABVIs and PAs are not specific to TBM but can be of tuberculous origin. Therefore, they do have a diagnostic value in the identification of TB in human osteoarchaeological material, especially when they simultaneously occur with other probable TB-related lesions. Their prudent utilization provides paleopathologists with a stronger basis for diagnosing TB and consequently, a more sensitive means of assessing TB frequency in past human populations.

An Investigation of Micro-CT Analysis of Bone as a New Diagnostic Method for Paleopathological Cases of Osteomalacia.

OBJECTIVE: This paper looks to broaden the methodological possibilities for diagnosing osteomalacia in archaeological bone using micro-CT analysis. Increasing the identification of osteomalacia in paleopathology will provide support for important interpretive frameworks. MATERIALS: Nine embedded and two unembedded rib fragments were sourced from St. Martin's Birmingham and Ancaster, UK, and Lisieux Michelet, France. Of the 11 samples, nine were previously confirmed as osteomalacic, and presented with varying levels of diagenesis and two were non-osteomalacic controls, one of which exhibits diagenetic change. METHODS: Micro-CT, backscattered scanning electron microscopy, and light microscopy were employed. Micro-CT images were evaluated for osteomalacic features using corresponding microscopic images. RESULTS: Micro-CT images from osteomalacic samples demonstrated the presence of defective mineralization adjacent to cement lines, areas of incomplete mineralization, and resorptive bays/borders, three key diagnostic features of osteomalacia. Diagenetic change was also detectable in micro-CT images, but did not prevent the diagnosis of osteomalacia. CONCLUSIONS: Micro-CT analysis is a non-destructive method capable of providing microstructural images of osteomalacic features in embedded and unembedded samples. When enough of these features are present, micro-CT images are capable of confirming a diagnosis of osteomalacia. SIGNIFICANCE: Vitamin D deficiency has important health consequences which operate throughout the life course. Increasing the ability to detect cases of vitamin D deficiency provides researchers with a greater understanding of health and disease in past communities. LIMITATIONS: Only adult rib samples were used. SUGGESTIONS FOR FURTHER RESEARCH: Paleopathologists should look to test the utility of micro-CT analysis in diagnosing active rickets in subadult individuals.