Abstract

Paleomicrobiological investigations of a 14th-century coprolite found inside a barrel in Namur, Belgium were done using microscopy, a culture-dependent approach and metagenomics. Results were confirmed by ad hoc PCR – sequencing. Investigations yielded evidence for flora from ancient environment preserved inside the coprolite, indicated by microscopic observation of amoebal cysts, plant fibers, seeds, pollens and mold remains. Seventeen different bacterial species were cultured from the coprolite, mixing organisms known to originate from the environment and organisms known to be gut inhabitants. Metagenomic analyses yielded 107,470 reads, of which known sequences (31.9%) comprised 98.98% bacterial, 0.52% eukaryotic, 0.44% archaeal and 0.06% viral assigned reads. Most abundant bacterial phyla were Proteobacteria, Gemmatimonadetes, Actinobacteria and Bacteroidetes. The 16 S rRNA gene dataset yielded 132,000 trimmed reads and 673 Operational Taxonomic Units. Most abundant bacterial phyla observed in the 16 S rRNA gene dataset belonged to Proteobacteria, Firmicutes, Actinobacteria and Chlamydia. The Namur coprolite yielded typical gut microbiota inhabitants, intestinal parasites Trichuris and Ascaris and systemic pathogens Bartonella and Bordetella. This study adds knowledge to gut microbiota in medieval times.

Highlights

  • Human paleomicrobiology, the quest for microbes in ancient specimens derived from humans, mainly relied on the investigations of old bone and dental pulp specimens [1]

  • In 2007, the specimen was transferred to the laboratory for paleomicrobiology of Aix-Marseille University, Marseille, France for further investigations

  • After aseptically peeling of its external portion, the inner portion of the coprolite was re-suspended in sterile Page’s amoeba saline medium (PAS) and microscopic observations revealed the presence of several eggs (Figure S1 in File S1)

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Summary

Introduction

The quest for microbes in ancient specimens derived from humans, mainly relied on the investigations of old bone and dental pulp specimens [1]. Less than 20 coprolites and ancient colon content samples collected from six American and two European archeological sites have been investigated using large-scale sequencing and PCR-based analyses These investigations yielded data about ancient gut microbiota, indicating that parts of the digestive flora were preserved in such specimens [2,4,5,7,8,9,10]. A recent study indicated that coprolites exhibited more similarities between each other, and with stools from modern rural communities, than with stools coming from modern cosmopolitan communities These findings support that modern lifestyle may participate to changes in the composition of the human gut flora [4]

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