Palmer, E. I. | bioRxiv Science

bioRxiv2026

Radiographic assessment of bone maturation as a tool for age estimation in common dolphins (Delphinus delphis)

We present the first radiographic ageing framework for common dolphins (Delphinus delphis), based on ossification and epiphyseal fusion patterns in the pectoral flipper, demonstrating higher reliability for chronological age estimation than currently available epigenetic approaches for this species. Using individuals of known dental age, we calibrated two modelling approaches to predict dental age from radiographic bone scores: 1) a univariate polynomial regression using a total bone score (sum of 16 scores across all assessed flipper bones), and 2) a multivariate canonical analysis of principal coordinates (CAP) incorporating 16 individual bone-score variables. Both approaches successfully predicted dental age from skeletal ossification patterns. For an age range of 0 to 24 years, polynomial regression demonstrated high predictive accuracy with median absolute errors (MAEs) of 1.25 years in females (Spearmans {rho} = 0.93, R{superscript 2} = 0.90) and 1.08 years in males ({rho} = 0.95, R{superscript 2} = 0.86). The CAP model yielded MAEs of 1.35 years in females ({rho} = 0.90, R{superscript 2} = 0.85) and 1.80 years in males ({rho} = 0.94, R{superscript 2} = 0.84). Notably, both radiographic bone ageing models achieved equal or lower median absolute errors and higher coefficients of determination than a recently developed epigenetic clock for common dolphins derived from the same population (MAE = 1.80, Pearsons correlation (r) = 0.91, R{superscript 2} = 0.82). When applying the bone ageing models to individuals of unknown dental age, both models produced age estimates consistent with expected life-history stages (foetus, neonate, juvenile, subadult, adult), although accuracy declined in dolphins above 20 years, likely as a consequence of subtle age-related variation in skeletal changes in this species. Radiographic ageing provides an accurate non-invasive tool for demographic assessment to support conservation management of common dolphins.

zoology↗

bioRxiv2025

Dental aging offers new insights to the first epigenetic clock for common dolphins (Delphinus delphis)

Determining exact age in wild odontocetes is essential for understanding population dynamics, survival, and reproduction, yet remains logistically challenging. While epigenetic aging is emerging as a valuable approach, only nine species-specific clocks currently exist. Most have been calibrated using captive known-age animals or well-studied wild populations. Only two previous studies have used dental ages from stranded or bycaught individuals. This is due to concerns that dental age inaccuracies, especially in older animals, may affect epigenetic clock performance. To explore this, we developed the first species-specific epigenetic clock for common dolphins (Delphinus delphis), analysing DNA methylation at 37,492 cytosine-phosphate-guanine sites in skin samples from stranded and bycaught dolphins with estimated dental ages. Elastic net models with Leave-One-Out Cross-Validation were applied to three subsets: the relaxed subset (all individuals; n = 75, median absolute error (MAE) = 2.02, r = 0.81, R2 = 0.66), the strict subset (excluding individuals with minimum dental age estimates only; n = 73, MAE = 2.29, r = 0.81, R2 = 0.66), and the restricted subset (excluding outliers with prediction errors > 6 years; n = 63, MAE = 1.80, r = 0.91, R2 = 0.82) to compare performance. Our models consistently underestimated the age of dolphins >16 years, even when minimum dental ages were applied, suggesting absolute errors between dental and epigenetic estimates unlikely reflect dental aging error. Additionally, post-mortem decomposition condition code (DCC 1 to 3) did not affect age prediction, signalling promise for future epigenetic clocks calibrated with strandings and bycaught individuals.

genetics↗