Integration of age-at-death and novel peptide sex estimation reveals cattle culling practices at Yorkshire Iron Age shrine site

Amelogenin peptide sex estimation is growing in bioarchaeological applications, with new studies focusing on developing rapid and inexpensive methods to enable sex estimations of larger numbers of samples. The main focus of these studies is hominin remains, with very few publications implementing amelogenin peptide analysis for faunal sex estimations, and no publications successfully employing high-throughput methods for large numbers of faunal remains. Here, alterations were made to previously reported TIMS-TOF (trapped ion mobility-time of flight) mass spectrometry methods (Blacka et al., 2025) to improve the applicability of amelogenin sex estimation to cattle remains. This improved method was implemented to analyse 40 teeth from cattle skulls excavated from a shrine/sanctuary associated with a Late Bronze Age/Early Iron Age ring fort at Kipling House Farm, near Middleton-on-the-Wolds, East Riding of Yorkshire, UK, enabling insights into the potential herd structure of the deposition. The rapid amelogenin method results agreed with the ancient DNA (aDNA) sex assignments for all the samples where both were undertaken, but the peptide analysis enabled extension of the dataset, doubling the number of sex estimations. When combined with age-at-death estimates, sex biases were revealed in the age structure, indicating a standard herd management. This therefore suggests that these cattle were not preferentially selected to be sacrificed, but instead may have been animals whose slaughter reflected normal culling practices. This study highlights the value in high throughput amelogenin sex estimation methods for achieving a deeper understanding of zooarchaeological remains. • Previously published rapid amelogenin sex estimation methods have been adapted to improve data quality for cattle. • Rapid palaeoproteomic amelogenin sex estimation was employed for 40 Iron Age cattle skulls. • aDNA sex assignments of 20 of the skulls were in full agreement with palaeoproteomic sex estimations. • Correlation of age-at-death and sex estimations reveal patterns possibly indicative of husbandry slaughter practices.