# Decoding past microbial life and antibiotic resistance in İnonü Cave’s archaeological soil

**Authors:** Sukran Ozturk, F.Gülden Ekmen, Hamza Ekmen, Esra Mine Ünal, Ayşegül Er, Emre Keskin, Benjamin Stanley Arbuckle, Peter F. Biehl, Peter F. Biehl, Peter F. Biehl, Peter F. Biehl

PMC · DOI: 10.1371/journal.pone.0326358 · PLOS One · 2025-07-31

## TL;DR

This study explores ancient microbial life and antibiotic resistance in soil from a Turkish cave, linking human history to changes in microbial communities.

## Contribution

The study identifies antibiotic-resistance genes in ancient soil samples, revealing long-term human impact on microbial evolution.

## Key findings

- Prominent bacterial groups like Acidobacteriota and Proteobacteria were found in ancient soil samples.
- Antibiotic resistance genes tetA, intl1, and OXA58 were detected in different historical periods.
- Human activities and environmental factors are linked to the persistence of resistance genes over time.

## Abstract

This study, which bridges the disciplines of archaeology and microbiology, examines the ancient bacterial communities and antibiotic-resistance genes in soil samples collected from İnönü Cave in Zonguldak, Turkiye. Our aim is to provide a comprehensive understanding of historical human activities and their influence on microbial communities. Soil samples were gathered from four distinct cultural levels from the Chalcolithic Age to the Early Iron Age. The microbial communities were characterized, and antibiotic-resistance genes were identified using high-throughput sequencing of 16S rRNA genes and metagenomic studies. This interdisciplinary approach not only enriches our understanding of ancient microbial communities but also opens up new avenues for research and collaboration. The results of our study showed a wide range of microorganisms, including prominent bacterial groups such as Acidobacteriota, Actinobacteriota, Bacteroidota, Chloroflexi, Cyanobacteria, Firmicutes, Myxococcota, and Proteobacteria. The study identified the presence of the tetracycline resistance gene tetA in Chalcolithic samples, the class 1 integron intl1 in Early Bronze Age samples, and the oxacillinase gene OXA58 in Late Bronze Age samples. These findings underscore the long-term impact of human activities on microbial communities, as antibiotic-resistance genes have been present and have remained over various historical periods, perhaps influenced by both human activities and environmental variables. This knowledge is crucial for understanding the resilience and adaptability of microbial communities in the face of human-induced changes. The coexistence of these resistance genes and alterations in the microbial population suggest substantial connections between human activities and soil microbiota. This study, which draws on the fields of archaeology, microbiology, and environmental science, offers valuable insights into the ancient microbial ecology and underscores the enduring presence of antibiotic resistance. It emphasizes the necessity of a comprehensive, interdisciplinary approach, spanning multiple fields, to comprehend microbial communities’ evolution and resistance mechanisms in archaeological settings.

## Linked entities

- **Genes:** tet(A) (tetracycline efflux MFS transporter Tet(A)) [NCBI Gene 33941499], itln1 (intelectin 1) [NCBI Gene 767684]

## Full-text entities

- **Genes:** TAF1A (TATA-box binding protein associated factor, RNA polymerase I subunit A) [NCBI Gene 9015] {aka MGC:17061, RAFI48, SL1, TAFI48}, ITLN1 (intelectin 1) [NCBI Gene 55600] {aka HL-1, HL1, INTL, ITLN, LFR, hIntL}, MMP10 (matrix metallopeptidase 10) [NCBI Gene 4319] {aka SL-2, STMY2}
- **Diseases:** neuralgia (MESH:D009437), diseases (MESH:D004194), rheumatism (MESH:D012216), gastrointestinal diseases (MESH:D005767), nitrogen pollution (MESH:D007222), antibiotic (MESH:D004761), poisonings (MESH:D011041), ORCID iD (MESH:C535742), infections (MESH:D007239), infectious diseases (MESH:D003141), Black Death (MESH:D010930)
- **Chemicals:** calcium carbonate (MESH:D002119), tetracycline (MESH:D013752), cephalosporins (MESH:D002511), activated charcoal (MESH:D002606), chlorophyllide (MESH:D002735), Aniline (MESH:C023650), carbapenem (MESH:D015780), chlorophyll (MESH:D002734), metal (MESH:D008670), nitric oxide (MESH:D009569), sulfur (MESH:D013455), MgCl2 (MESH:D015636), Agarose (MESH:D012685), gold (MESH:D006046), copper (MESH:D003300), C14 carbon (-), carbon (MESH:D002244), water (MESH:D014867), nitrate (MESH:D009566), nitrite (MESH:D009573), tylosin (MESH:D015645), polysaccharides (MESH:D011134), Iron (MESH:D007501), nitrogen (MESH:D009584)
- **Species:** Capreolus capreolus (Western roe deer, species) [taxon 9858], Terriglobia (class) [taxon 204432], Clostridium (genus) [taxon 1485], Bos taurus (bovine, species) [taxon 9913], uncultured cyanobacterium (species) [taxon 1211], Cervidae (deer, family) [taxon 9850], Ovis aries (domestic sheep, species) [taxon 9940], Acidobacteriota (phylum) [taxon 57723], Bacillus (genus) [taxon 55087], Staphylococcus aureus (species) [taxon 1280], Lactobacillus (genus) [taxon 1578], Cyanobacteriota (blue-green algae, phylum) [taxon 1117], Nitrospirota (phylum) [taxon 40117], Sus scrofa (pig, species) [taxon 9823], Equus caballus (domestic horse, species) [taxon 9796], Cervus elaphus (red deer, species) [taxon 9860], Bacteria Latreille et al. 1825 (Bacteria stick insect, genus) [taxon 629395], Homo sapiens (human, species) [taxon 9606], Actinomycetota (actinobacteria, phylum) [taxon 201174], Capra hircus (domestic goat, species) [taxon 9925], Pseudomonadota (proteobacteria, phylum) [taxon 1224], Bacillota (clostridial firmicutes, phylum) [taxon 1239], Bacteroidota (Bacteroides-Cytophaga-Flexibacter group, phylum) [taxon 976]

## Full text

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## Figures

8 figures with captions in the complete paper: https://tomesphere.com/paper/PMC12312911/full.md

## References

67 references — full list in the complete paper: https://tomesphere.com/paper/PMC12312911/full.md

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Source: https://tomesphere.com/paper/PMC12312911