Biodiversity loss, a largely anthropogenically driven phenomenon, represents a monumental challenge for both the natural and human world that has serious repercussions. Animal Ambassadors are used by zoological parks and similar institutions to provide close encounter experiences to visitors designed to enhance pro-conservation sentiments. A scoping review was undertaken to assess the animal ambassador effect on human pro-environmental knowledge, attitudes and behaviours, following the PRISMA-ScR guidelines. A search was conducted using the databases Medline, PsycInfo, Scopus and Web of Science, and a grey literature search was undertaken using the ProQuest Dissertation and Theses Global Database, along with the search engine Google (advanced search) and Google Scholar (advanced search). The results demonstrate some acute gains in pro-environmental sentiments; however, longitudinal studies indicate these are largely lost overtime. The importance of the animal in animal ambassador programs is also questioned, with other alternative educational experiences discussed.