# New insights from the application of ZooMS to Late Pleistocene fauna from Grotta di Castelcivita, southern Italy

## TL;DR

This study uses ZooMS to identify fragmented bones from a Paleolithic cave in Italy, revealing new insights into the faunal assemblage and subsistence patterns.

## Contribution

The study demonstrates the effectiveness of ZooMS in identifying Pleistocene faunal remains in southern Italy, revealing previously unknown species and abundance patterns.

## Key findings

- ZooMS identified a significant increase in equids and bovids during the Uluzzian and Protoaurignacian periods.
- New identifications of rhinoceros, bear, and canids were found in deeper layers of the cave.
- High collagen preservation in the bones suggests potential for further biomolecular studies in the region.

## Abstract

The Middle to Upper Paleolithic cave site of Grotta di Castelcivita (Campania, Southern Italy) contains a key archaeological sequence exhibiting Late Mousterian occupation followed by the Uluzzian techno-complex and an Aurignacian sequence (Protoaurignacian and Early Aurignacian). Abundant faunal remains are found throughout the sequence with variations in taxa present in each period. Previous studies of the morphologically identifiable faunal remains have provided valuable information on species abundance and diversity to reconstruct subsistence behaviour. However, like in many Pleistocene sequences, much of the faunal assemblage is fragmented and unidentifiable. Here we focus on these unidentified fragmentary bones to add greater dimension to the observed patterns. The application of collagen peptide mass fingerprinting (or Zooarchaeology by Mass Spectrometry; ZooMS) on 1263 unidentified bones revealed distinct changes in ZooMS NISP values in the Uluzzian and Protoaurignacian periods where equids and bovids nearly doubled in quantity compared to the original morphologically identified macrofaunal assemblage. New ZooMS identifications of rhinoceros, bear, and canids were made in layers deeper than previous recorded zooarchaeological analyses, extending the presence of these taxa at the site. The unexpectedly high level of collagen preservation in the bones from the cave confirms the potential for further applications of biomolecular approaches to Pleistocene bones from southern Italy.

The online version contains supplementary material available at 10.1038/s41598-025-11355-6.