Tentative indicators of malaria in archaeological skeletal samples, a pilot study testing different methods.

OBJECTIVE: This study attempts to integrate multiple methods to investigate the presence of malaria in human skeletal samples from an archaeological context. MATERIALS: 33 well preserved human remains originating from a 17th-century archaeological site in southeastern Romania. METHODS: The human bone samples were analyzed using rapid diagnostic tests for malaria antigens and PCR amplification of Plasmodium falciparum apical membrane antigen 1. A preliminary test was performed to identify and briefly characterize the presence of hemozoin using a combination of TEM imaging and diffraction. RESULTS: The rapid diagnostic tests indicated that more than half of the examined samples were positive for Plasmodium antigens, but no traces of the parasites' genetic material were detected despite repeated attempts. The TEM images indicated that hemozoin might be a promising diagnostic marker of malaria in ancient bones. CONCLUSIONS: The indisputable identification of malaria in the analyzed archaeological population was not possible as none of the applied methodological strategies turned out to be straightforward. SIGNIFICANCE: This study reinforces the intricacy and limitations of unequivocally identifying malaria in past populations and sets the stage for future studies on such life-threatening infectious disease in a geographical space, which is currently underrepresented in the bioarchaeological literature. LIMITATIONS: The low sample size and the lack of consistency across all assays hindered understanding the role of malaria in the studied population. SUGGESTIONS FOR FURTHER RESEARCH: Further thorough multidisciplinary approaches on malaria detection in ancient settlements would be appropriate to inform our knowledge of its origins, frequency, and pathogen changes over centuries.

Mitochondrial ancestry of medieval individuals carelessly interred in a multiple burial from southeastern Romania.

The historical province of Dobruja, located in southeastern Romania, has experienced intense human population movement, invasions, and conflictual episodes during the Middle Ages, being an important intersection point between Asia and Europe. The most informative source of maternal population histories is the complete mitochondrial genome of archaeological specimens, but currently, there is insufficient ancient DNA data available for the medieval period in this geographical region to complement the archaeological findings. In this study, we reconstructed, by using Next Generation Sequencing, the entire mitochondrial genomes (mitogenomes) of six medieval individuals neglectfully buried in a multiple burial from Capidava necropolis (Dobruja), some presenting signs of a violent death. Six distinct maternal lineages (H11a1, U4d2, J1c15, U6a1a1, T2b, and N1a3a) with different phylogenetic background were identified, pointing out the heterogeneous genetic aspect of the analyzed medieval group. Using population genetic analysis based on high-resolution mitochondrial data, we inferred the genetic affinities of the available medieval dataset from Capidava to other ancient Eurasian populations. The genetic data were integrated with the archaeological and anthropological information in order to sketch a small, local piece of the mosaic that is the image of medieval European population history.

Maternal DNA lineages at the gate of Europe in the 10th century AD.

Given the paucity of archaeogenetic data available for medieval European populations in comparison to other historical periods, the genetic landscape of this age appears as a puzzle of dispersed, small, known pieces. In particular, Southeastern Europe has been scarcely investigated to date. In this paper, we report the study of mitochondrial DNA in 10th century AD human samples from Capidava necropolis, located in Dobruja (Southeastern Romania, Southeastern Europe). This geographical region is particularly interesting because of the extensive population flux following diverse migration routes, and the complex interactions between distinct population groups during the medieval period. We successfully amplified and typed the mitochondrial control region of 10 individuals. For five of them, we also reconstructed the complete mitochondrial genomes using hybridization-based DNA capture combined with Next Generation Sequencing. We have portrayed the genetic structure of the Capidava medieval population, represented by 10 individuals displaying 8 haplotypes (U5a1c2a, V1a, R0a2'3, H1, U3a, N9a9, H5e1a1, and H13a1a3). Remarkable for this site is the presence of both Central Asiatic (N9a) and common European mtDNA haplotypes, establishing Capidava as a point of convergence between East and West. The distribution of mtDNA lineages in the necropolis highlighted the existence of two groups of two individuals with close maternal relationships as they share the same haplotypes. We also sketch, using comparative statistical and population genetic analyses, the genetic relationships between the investigated dataset and other medieval and modern Eurasian populations.