Age is an important ecological tool in wildlife conservation. However, it is difficult to estimate in most animals, including felines — most of whom are endangered. Here, we developed the first DNA methylation-based age-estimation technique — as an alternative to current age-estimation methods — for two feline species that share a relatively long genetic distance with each other: domestic cat (Felis catus; 79 blood samples) and an endangered Panthera, the snow leopard (Panthera uncia; 11 blood samples). We measured the methylation rates of two gene regions using methylation-sensitive high-resolution melting (MS-HRM). Domestic cat age was estimated with a mean absolute deviation (MAD) of 3.83 years. Health conditions influenced accuracy of the model. Specifically, the models built on cats with chronic kidney disease (CKD) had lower accuracy than those built on healthy cats. The snow leopard-specific model (i.e. the model that resets the model settings for snow leopards) had a better accuracy (MAD = 2.10 years) than that obtained on using the domestic cat model directly. This implies that our markers could be utilised across species, although changing the model settings when targeting different species could lead to better estimation accuracy. The snow leopard-specific model also successfully distinguished between sexually immature and mature individuals.

The cat data and the snow leopard data csv files are the raw data used in the analysis. The R script could be referred to in both the R markdown file and the pdf file. Please refer to Readme file to see the detailed explanation for the datasets. Funding provided by: Japan Society for the Promotion of ScienceCrossref Funder Registry ID: http://dx.doi.org/10.13039/100014423Award Number: 17K19426Funding provided by: Japan Society for the Promotion of ScienceCrossref Funder Registry ID: http://dx.doi.org/10.13039/501100001691Award Number: JP20H03008Funding provided by: Japan Society for the Promotion of ScienceCrossref Funder Registry ID: http://dx.doi.org/10.13039/501100001691Award Number: JPJSBP120219921Funding provided by: Japan Society for the Promotion of ScienceCrossref Funder Registry ID: http://dx.doi.org/10.13039/501100001691Award Number: JPJSBP120209915Funding provided by: Environmental Restoration and Conservation AgencyCrossref Funder Registry ID: http://dx.doi.org/10.13039/100014423Award Number: JPMEERF20214001