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Recent research has emphasised the importance of understanding reproductive biology and physiology, behavior, pain management, internal medicine, pathogen prevalence, dental health, and overall wellbeing—all areas critical to the conservation and sustainable management of donkey and mule populations. Studies have also addressed anatomical and pharmacokinetic differences between horses and donkeys, highlighting their unique needs.Assisted reproductive techniques (ARTs), such as artificial insemination (AI), sperm cryopreservation, and embryo transfer (ET), which are explored in this Special Issue, play a pivotal role in preserving genetic resources.Animal genetic resources are priceless heritage, making it crucial to safeguard genetic diversity across species, breeds, varieties, and native ecotypes. Many donkey breeds are currently classified as endangered and face significant challenges, including inbreeding, poor reproductive management, advanced age, and low rates of reproduction.This Special Issue brings together contributions from researchers worldwide, showcasing the latest advancements in donkey and mule research. It provides an updated overview of the primary conditions affecting the health of these species, offering valuable insights into the development of new strategies and paving the way for future studies.

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