Mpox, known before now as monkeypox, is a novel zoonotic disease that may infect both humans and animals. It usually spreads by direct contact with bodily fluids, lesion material, or fomites (such as contaminated linens) and prolonged face-to-face contact. Mpox can spread to laboratory animals in a research laboratory through either a general outbreak or during procedures. Animal models are essential as we learn more about how infections occur and how to treat illnesses in both humans and animals which require laboratory procedures with mpox virus. As drug-resistant organisms proliferate, bioterrorism becomes a greater danger, international trade and travel expand, and the number of newly developing infectious illnesses grows, the use of animals in infectious disease research has increased. Animal research offers a solid basis for clinical trials used in standard medication development.Animal studies must be planned to produce conclusive effectiveness evidence that is robust, rigorous, and repeatable per the animal rule so that the agency may depend on the results when making regulatory decisions.There is a need to understand the transmission risk and biosafety protocol to be in place in laboratory settings to prevent an outbreak and probably contain an outbreak. Hence, this review discussed the overview of mpox in animal research, biosafety protocols in mpox research, laboratory waste management methods, legal and ethical considerations in mpox research, challenges in carrying out mpox research and future directions.