We assessed wildlife predation on livestock and poultry and the implications for predator conservation in the most important rainforest areas of south-east Mexico. We estimated the economic impact of predation and identified variables correlated to predation frequency in four communities adjacent to Calakmul and Montes Azules Biosphere Reserves. We did participant observation and interviews with 161 residents during January–September 2010. We recorded 2,861 deaths of livestock and poultry from predation by wild carnivores over the previous 3 years. Opossums (n = 677 predation events), raptors (n = 676) and ocelots (n = 539) were responsible for the highest numbers of killings. Poultry constituted the main target of wild predators (n = 2,548 deaths). Sheep (n = 201 events) and cattle (167) were the most affected by large predators such as jaguar Panthera onca and puma Puma concolor. Economic losses from predation were estimated to be USD 55,600 over the 3 years in the four communities. Predation frequency was positively correlated with livestock numbers (r2 = 0.87, P < 0.05) but negatively associated with wild prey abundance (r2 = 0.96, P < 0.05). Better husbandry practices focused on the safety and location of livestock enclosures and on reducing overabundance of dogs and regulating hunting on wild prey in rural areas may help mitigating conflict between wild predators and residents in the rainforests of south-east Mexico.