DNA methylation levels in candidate genes associated with chronological age in mammals are not conserved in a long-lived seabird

<div><p>Most seabirds do not have any outward identifiers of their chronological age, so estimation of seabird population age structure generally requires expensive, long-term banding studies. We investigated the potential to use a molecular age biomarker to estimate age in short-tailed shearwaters (<i>Ardenna tenuirostris</i>). We quantified DNA methylation in several <i>A</i>. <i>tenuirostris</i> genes that have shown age-related methylation changes in mammals. In birds ranging from chicks to 21 years of age, bisulphite treated blood and feather DNA was sequenced and methylation levels analysed in 67 CpG sites in 13 target gene regions. From blood samples, five of the top relationships with age were identified in <i>KCNC3</i> loci (CpG66: <i>R</i>² = 0.325, <i>p</i> = 0.019). In feather samples <i>ELOVL2</i> (CpG42: <i>R</i>² = 0.285, <i>p</i> = 0.00048) and <i>EDARADD</i> (CpG46: <i>R</i>² = 0.168, <i>p</i> = 0.0067) were also weakly correlated with age. However, the majority of markers had no clear association with age (of 131 comparisons only 12 had a p-value < 0.05) and statistical analysis using a penalised lasso approach did not produce an accurate ageing model. Our data indicate that some age-related signatures identified in orthologous mammalian genes are not conserved in the long-lived short tailed shearwater. Alternative molecular approaches will be required to identify a reliable biomarker of chronological age in these seabirds.</p>