Traditional Ecological Knowledge (TEK) and folklore are repositories of large amounts of information about the natural world. Ideas, perceptions and empirical data held by human communities regarding local species are important sources which enable new scientific discoveries to be made, as well as offering the potential to solve a number of conservation problems. We documented the gecko-related folklore and TEK of the people of southern Portugal, with the particular aim of understanding the main ideas relating to gecko biology and ecology. Our results suggest that local knowledge of gecko ecology and biology is both accurate and relevant. As a result of information provided by local inhabitants, knowledge of the current geographic distribution of Hemidactylus turcicus was expanded, with its presence reported in nine new locations. It was also discovered that locals still have some misconceptions of geckos as poisonous and carriers of dermatological diseases. The presence of these ideas has led the population to a fear of and aversion to geckos, resulting in direct persecution being one of the major conservation problems facing these animals. It is essential, from both a scientific and conservationist perspective, to understand the knowledge and perceptions that people have towards the animals, since, only then, may hitherto unrecognized pertinent information and conservation problems be detected and resolved.