SUMMARYAnaplasmataceae agents comprise obligate intracellular bacteria that can cause disease in humans and animals. Between August 2013 and March 2015, 31Nasua nasua(coati), 78Cerdocyon thous(crab-eating fox), sevenLeopardus pardalis(ocelot), 110 wild rodents, 30 marsupials, and 42 dogs were sampled in the Pantanal wetland, Brazil. In addition, ectoparasites found parasitizing the animals were collected and identified. The present work aimed to investigate the occurrence of Anaplasmataceae agents in wild mammals, domestic dogs and ectoparasites, by molecular and serological techniques. Overall, 14 (17·9%)C. thous, seven (16·6%) dogs and one (3·2%)N. nasuawere seroreactive toEhrlichia canis. Nine dogs, twoC. thous,oneN. nasua, eight wild rodents, five marsupials, eightAmblyomma sculptum,fourAmblyomma parvum,13A. sculptumnymphal pools, twoAmblyommalarvae pools and onePolygenis (Polygenis) bohlsi bohlsiflea pool were positive forEhrlichiaspp. closely related toE. canis. SevenN. nasua, two dogs, oneC. thous,oneL. pardalis, four wild rodents, three marsupials, 15A. sculptum, twoAmblyomma ovale,twoA. parvumand oneAmblyommaspp. larval pools were positive forAnaplasmaspp. closely related toA. phagocytophilumorA. bovis. The present study provided evidence that wild animals from Brazilian Pantanal are exposed to Anaplasmataceae agents.