The Western European hedgehog (Erinaceus europaeus) is an insectivorous mammal with a wide geographic distribution. Owing mostly to climate changes and anthropogenic pressures, a considerable number of hedgehogs now live in urban areas close to humans, where they are exposed to contaminants and biological agents that may result in disease with the correspondent hematological and biochemical alterations. Hedgehogs can work as bioindicators to environmental pollution and host multiple zoonotic agents, making them relevant for One Health studies. Thus, it is essential to deepen the knowledge on this species and calculate reference intervals for the usual hematological and biochemical parameters. This would make it possible to recognize the “normal” and identify the “disease”. In this study, some significant differences were evident, especially when comparing age groups (juveniles versus adults), showing the relevance of further investigations in this species.