Simple SummaryConsiderable evidence suggests that skin microbiota communities are critical for maintaining health and skin homeostasis. However, to date, studies on the skin microorganisms of donkeys are surprisingly rare. Therefore, the present study investigated the dynamic changes in commensal microbial communities (bacteria and fungal composition) on the skins of healthy donkeys throughout the growing period. Alpha and beta diversity analysis showed that bacterial and fungal communities on the skin display obvious age-related variations, especially at 1 month and 48 months of the growing period. Notably, these changes are also affected by the living environment. Our data could provide insights into microbial involvement in skin disorders and give useful information for improving health management and maintaining the healthy skin of Dezhou donkeys.AbstractConsiderable evidence suggests that the skin microbiota is not only important and complex in humans and other mammals but also critical for maintaining health and skin homeostasis. To date, studies on the skin microorganisms of donkeys are surprisingly rare. To investigate the dynamic changes in commensal microbial communities on the skins of healthy donkeys throughout the growing period, skin and soil samples were collected from 30 healthy Dezhou donkeys (ranging from 1, 6, 12, 24 to 48 months of age) and their corresponding breeding sheds on the farm. All samples were analysed for high-throughput sequencing of the 16S rRNA and ITS to characterize the skin microbiota of healthy donkeys and compare the differences in skin microbiota among donkeys of different ages. There were notable differences in the proportions of various genera (including bacteria and fungi) between dorsal and abdominal skin with increasing age. The comparison of the skin microbial communities among these groups revealed thatStaphylococcuswas mainly enriched in the early growing stage (1 and 6 months), while the relative abundance ofStreptococcuswas higher in both the 1- and 48-month-old age groups. Moreover, some bacteria and commensal fungi, such asStaphylococcusandTrichosporon, were found to be positively correlated between the skin and the environment. This is the first study to investigate the dynamic changes in skin microbiota diversity and composition in donkeys of different ages and at different sites of the body. Furthermore, this study provides insights into the dynamic alterations in skin microbes during a donkey’s growth and characterizes the profiles of bacterial and fungal communities across a donkey’s body regions (dorsal and abdomen).