Abstract

Photogrammetry and cascading microscopy investigations of dental pulp specimens collected from 2,000-year-old individuals buried in a Roman necropolis in Besançon, France, revealed unprecedented preserved tissular and cellular morphology. Photogrammetry yielded 3-D images of the smallest archaeological human remains ever recovered. Optical microscopy examinations after standard haematoxylin-phloxine-saffron staining and anti-glycophorin A immunohistochemistry exposed dental pulp cells, in addition erythrocytes were visualised by electron microscopy, which indicated the ancient dental pulp trapped a blood drop. Fluorescence in situ hybridisation applied on red blood cells revealed the louse-borne pathogen Bartonella quintana, a finding confirmed by polymerase chain reaction assays. Through paleohistology and paleocytology, we demonstrate that the ancient dental pulp preserved intact blood cells at the time of the individual’s death, offering an unprecedented opportunity to engage in direct and indirect tests to diagnose pathogens in ancient buried individuals.

Highlights

  • Photogrammetry and cascading microscopy investigations of dental pulp specimens collected from 2,000-year-old individuals buried in a Roman necropolis in Besançon, France, revealed unprecedented preserved tissular and cellular morphology

  • Using paleocytology, we show that ancient dental pulp can preserve blood cells exhibiting unanticipated and perfectly conserved morphology

  • Microscopic detection of B. quintana was confirmed by two different quantitative real-time PCR (qPCR) assays, including negative controls and no positive controls

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Summary

Introduction

Photogrammetry and cascading microscopy investigations of dental pulp specimens collected from 2,000-year-old individuals buried in a Roman necropolis in Besançon, France, revealed unprecedented preserved tissular and cellular morphology. Recent investigations unexpectedly revealed host peptides, including peptides derived from conjunctive dental pulp tissue and plasmatic peptides, such as coagulation factors and immunoglobulins[8]. These observations indicated that ancient dental pulp can preserve blood and its serum phase. Illustrating one outcome of this discovery in the field of paleomicrobiology, we performed microscopic-only detection of one ancient intracellular pathogen, Bartonella quintana[9], in a 2,000-year-old Roman dental pulp specimen obtained in France

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