Although hemoplasma infection has been widely described in animals, a few studies have been conducted involving human populations, mostly as case reports. This was a cross-sectional epidemiological study that accessed hemoplasma infection in individuals and dogs from ten Indigenous communities of southern and southeastern Brazil. A total of 23/644 (3.6%) individuals of ten Indigenous communities tested positive to hemoplasmas by quantitative polymerase chain reaction (qPCR) (cycle threshold
Ct ≤ 34.4), with 3/644 (0.5%) Mycoplasma haemocanis detection. In addition, 91/416 (21.9%) dogs were positive to hemoplasmas by qPCR, with 54/91 (59.3%) for M. haemocanis, 27/91 (29.7%) for Candidatus Mycoplasma haematoparvum, and 10/91 (11.0%) for both. Molecular diagnosis of M. haemocanis in Indigenous people herein may be consequence of daily close contact with dogs and different potential vectors. Although apparently healthy individuals, hemoplasma infection should be considered as differential diagnosis in likely overexposed populations, such as Indigenous individuals.