As the HIV/AIDS pandemic continues unabated, novel prophylactic strategy for the spread of HIV are urgently needed. Topical microbicides are designed to prevent transmission of HIV when applied vaginally or rectally. Although there are many microbicide candidates in the pipeline, animal models for evaluating their safety and efficacy are urgently needed. On the basis of comparing the non-primate small animal models and the non-human primate animal models in evaluating safety and efficacy of HIV microbicides, this review summarizes the major advantages and disadvantages of the relevant animal models. The suggested direction of research that would benefit the development of microbicides is also reviewed.