Ancient DNA and Paleoproteomic Analysis on Roman Imperial-Era Individuals from Histria, Romania

Ancient biomolecules have become an increasingly important part of archaeological investigations interested in understanding population movements and health. Despite their ability to elucidate historically-attested contexts of human mobility and interaction between different cultural groups, biomolecular techniques are still underutilized in certain historical and archaeological contexts. One such context is the Roman Imperial limes, or border zone, along the lower reaches of the Danube, which saw more than five hundred years of migration, conflict, and accommodation among a wide range of populations, from Mediterranean settlers to steppe pastoralists. In this region, more than a century of archaeological investigation has unearthed the remains of tens of thousands of Roman-era individuals. However, only a limited number of contexts have undergone biomolecular analyses. While applying biomolecular methods to these previously excavated human remains may offer an untapped reservoir of information, many were collected during a period when the standard precautions and protocols for ancient biomolecular researcher were not yet established. Because contamination is a major barrier for successfully recovering the ancient DNA and proteins from human remains, conducting a pilot study on a small dataset of human remains may minimize unnecessary destructive sampling. This study applies ancient DNA and paleoproteomic techniques to human remains from a Roman-period cemetery at Histria, a site located just south of the Danube at the edge of the Roman province of Moesia Inferior. These individuals were excavated between the 1940s and 1980s and have been held in storage since. Our results suggest that there is endogenous microbial and human DNA in the dental calculus and dentin samples, respectively. We also successfully recovered sex-specific amelogenin peptides in tooth enamel of three individuals, including a subadult. In conclusion, the results from this study are encouraging, signifying the feasibility of future aDNA and paleoproteomic research for this skeletal collection. Our findings not only open doors to gaining deeper insights into the individuals within this skeletal collection but also hold the promise of addressing broader anthropological inquiries, such as the impact of Roman annexation in this region.