This multi-sectoral zoonotic diseases outbreak investigation (ZDOBI) guideline is developed to provide guidance on coordination and zoonotic diseases outbreak investigation steps, aiming to effectively and efficiently detect, prevent and control the spread of disease outbreak. This guideline is the first version jointly developed by the Ethiopian Public Health Institute (EPHI), Ministry of Agriculture (MOA) and Ethiopian Environment Authority and Ethiopian Wildlife Conservation Authority Commission (EWCA), with support of development partners, meant to provide comprehensive guide for zoonotic disease outbreak investigation and initiation of quick response actions.