We conducted a survey questionnaire to assess and compare public perceptions and knowledge of jaguars Panthera onca and pumas Puma concolor among two stakeholder groups likely to come into conflict with these large carnivores: cattle ranchers living near protected areas and inhabitants of two villages located within the multiple-use forest of the Maya Biosphere Reserve, Guatemala. We tested whether stakeholder group (i.e. rancher or villager) or previous experience with livestock attacks by carnivores predicted a negative perception of jaguars and pumas. The odds of people fearing an attack on humans by these felids and of recognizing the ecological importance of these species was best explained by stakeholder group. Villagers exhibited a more negative perception towards these felids than did cattle ranchers. Our results highlight the importance of tailoring conservation programmes in high risk areas to the needs of specific stakeholder groups. Further research is needed to understand which factors (i.e. ecological settings, socio-economic differences) best explain these differences and whether these differences in perception vary within groups. Programmes that work with local people to assess the ecological importance of large predators and the true risks of carnivore attacks on livestock and humans along with low-cost conflict mitigation measures should be implemented to increase the level of tolerance towards these carnivores.